Glycosidase inhibitors and uses thereof

ABSTRACT

The invention provides compounds for inhibiting glycosidases, prodrugs of the compounds, and pharmaceutical compositions including the compounds or prodrugs of the compounds. The invention also provides methods of treating diseases and disorders related to deficiency or overexpression of O-GlcNAcase, accumulation or deficiency of O-GlcNAc.

FIELD OF THE INVENTION

This application relates to compounds which inhibit glycosidases anduses thereof.

BACKGROUND OF THE INVENTION

A wide range of cellular proteins, both nuclear and cytoplasmic, arepost-translationally modified by the addition of the monosaccharide2-acetamido-2-deoxy-β-D-glucopyranoside (β-N-acetylglucosamine) which isattached via an O-glycosidic linkage.¹ This modification is generallyreferred to as O-linked N-acetylglucosamine or O-GlcNAc. The enzymeresponsible for post-translationally linking β-N-acetylglucosamine(GlcNAc) to specific serine and threonine residues of numerousnucleocytoplasmic proteins is O-GlcNAc transferase (OGT).²⁻⁵ A secondenzyme, known as glycoprotein 2-acetamido-2-deoxy-β-D-glucopyranosidase(O-GlcNAcase)^(6,7) removes this post-translational modification toliberate proteins making the O-GlcNAc-modification a dynamic cycleoccurring several times during the lifetime of a protein.⁸

O-GlcNAc-modified proteins regulate a wide range of vital cellularfunctions including, for example, transcription,⁹⁻¹² proteasomaldegradation,¹³ and cellular signaling.¹⁴ O-GlcNAc is also found on manystructural proteins.¹⁵⁻¹⁷ For example, it has been found on a number ofcytoskeletal proteins, including neurofilament proteins,^(18,19)synapsins,^(6,20) synapsin-specific clathrin assembly protein AP-3,⁷ andankyrinG.¹⁴ O-GlcNAc modification has been found to be abundant in thebrain.^(21,22) It has also been found on proteins clearly implicated inthe etiology of several diseases including Alzheimer's disease (AD) andcancer.

For example, it is well established that AD and a number of relatedtauopathies including Downs' syndrome, Pick's disease, Niemann-Pick TypeC disease, and amyotrophic lateral sclerosis (ALS) are characterized, inpart, by the development of neurofibrillary tangles (NFTs). These NFTsare aggregates of paired helical filaments (PHFs) and are composed of anabnormal form of the cytoskeletal protein “tau”. Normally tau stabilizesa key cellular network of microtubules that is essential fordistributing proteins and nutrients within neurons. In AD patients,however, tau becomes hyperphosphorylated, disrupting its normalfunctions, forming PHFs and ultimately aggregating to form NFTs. Sixisoforms of tau are found in the human brain. In AD patients, all sixisoforms of tau are found in NFTs, and all are markedlyhyperphosphorylated.^(23,24) Tau in healthy brain tissue bears only 2 or3 phosphate groups, whereas those found in the brains of AD patientsbear, on average, 8 phosphate groups.^(25,26) A clear parallel betweenNFT levels in the brains of AD patients and the severity of dementiastrongly supports a key role for tau dysfunction in AD.²⁷⁻²⁹ The precisecauses of this hyperphosphorylation of tau remain elusive. Accordingly,considerable effort has been dedicated toward: a) elucidating themolecular physiological basis of tau hyperphosphorylation;³⁰ and b)identifying strategies that could limit tau hyperphosphorylation in thehope that these might halt, or even reverse, the progression ofAlzheimer's disease³¹⁻³⁴ Thus far, several lines of evidence suggestthat up-regulation of a number of kinases may be involved inhyperphosphorylation of tau,^(21,35,36) although very recently, analternative basis for this hyperphosphorylation has been advanced.²¹

In particular, it has emerged that phosphate levels of tau are regulatedby the levels of O-GlcNAc on tau. The presence of O-GlcNAc on tau hasstimulated studies that correlate 0-GlcNAc levels with tauphosphorylation levels. The interest in this field stems from theobservation that O-GlcNAc modification has been found to occur on manyproteins at amino acid residues that are also known to bephosphorylated.³⁷⁻³⁹ Consistent with this observation, it has been foundthat increases in phosphorylation levels result in decreased O-GlcNAclevels and conversely, increased O-GlcNAc levels correlate withdecreased phosphorylation levels.⁴⁰ This reciprocal relationship betweenO-GlcNAc and phosphorylation has been termed the “Yin-Yang hypothesis”⁴¹and has gained strong biochemical support by the discovery that theenzyme OGT⁴ forms a functional complex with phosphatases that act toremove phosphate groups from proteins.⁴² Like phosphorylation, O-GlcNAcis a dynamic modification that can be removed and reinstalled severaltimes during the lifespan of a protein. Suggestively, the gene encodingO-GlcNAcase has been mapped to a chromosomal locus that is linked toAD.^(7,43) Hyperphosphorylated tau in human AD brains has markedly lowerlevels of O-GlcNAc than are found in healthy human brains.²¹ It has beenshown that O-GlcNAc levels of soluble tau protein from human brainsaffected with AD are markedly lower than those from healthy brain.²¹Furthermore, PHF from diseased brain was suggested to lack completelyany O-GlcNAc modification whatsoever.²¹ The molecular basis of thishypoglycosylation of tau is not known, although it may stem fromincreased activity of kinases and/or dysfunction of one of the enzymesinvolved in processing O-GlcNAc. Supporting this latter view, in bothPC-12 neuronal cells and in brain tissue sections from mice, anonselective N-acetylglucosamindase inhibitor was used to increase tauO-GlcNAc levels, whereupon it was observed that phosphorylation levelsdecreased.²¹ The implication of these collective results is that bymaintaining healthy O-GlcNAc levels in AD patients, such as byinhibiting the action of O-GlcNAcase, one should be able to blockhyperphosphorylation of tau and all of the associated effects of tauhyperphosphorylation, including the formation of NFTs and downstreameffects. However, because the proper functioning of theβ-hexosaminidases is critical, any potential therapeutic interventionfor the treatment of AD that blocks the action of O-GlcNAcase would haveto avoid the concomitant inhibition of both hexosaminidases A and B.

Neurons do not store glucose and therefore the brain relies on glucosesupplied by blood to maintain its essential metabolic functions.Notably, it has been shown that within brain, glucose uptake andmetabolism decreases with aging.⁴⁴ Within the brains of AD patientsmarked decreases in glucose utilization occur and are thought to be apotential cause of neurodegeneration.⁴⁵ The basis for this decreasedglucose supply in AD brain⁴⁶⁻⁴⁸ is thought to stem from any of decreasedglucose transport,^(49,50) impaired insulin signaling,^(51,52) anddecreased blood flow.⁵³

In light of this impaired glucose metabolism, it is worth noting that ofall glucose entering into cells, 2-5% is shunted into the hexosaminebiosynthetic pathway, thereby regulating cellular concentrations of theend product of this pathway, uridine diphosphate-N-acetylglucosamine(UDP-GlcNAc).⁵⁴ UDP-GlcNAc is a substrate of the nucleocytoplasmicenzyme O-GlcNAc transferase (OGT),²⁻⁵ which acts to post-translationallyadd GlcNAc to specific serine and threonine residues of numerousnucleocytoplasmic proteins. OGT recognizes many of itssubstrates^(55,56) and binding partners^(42,57) through itstetratricopeptide repeat (TPR) domains.^(58,59) As described above,O-GlcNAcase^(6,7) removes this post-translational modification toliberate proteins making the O-GlcNAc-modification a dynamic cycleoccurring several times during the lifetime of a protein.⁸ O-GlcNAc hasbeen found in several proteins on known phosphorylationsites,^(10,38,39,60) including tau and neurofilaments.⁶¹ Additionally,OGT shows unusual kinetic behaviour making it exquisitely sensitive tointracellular UDP-GlcNAc substrate concentrations and therefore glucosesupply.⁴²

Consistent with the known properties of the hexosamine biosyntheticpathway, the enzymatic properties of OGT, and the reciprocalrelationship between O-GlcNAc and phosphorylation, it has been shownthat decreased glucose availability in brain leads to tauhyperphosphorylation.⁴⁵ Therefore the gradual impairment of glucosetransport and metabolism, whatever its causes, leads to decreasedO-GlcNAc and hyperphosphorylation of tau (and other proteins).Accordingly, the inhibition of O-GlcNAcase should compensate for the agerelated impairment of glucose metabolism within the brains of healthindividuals as well as patients suffering from AD or relatedneurodegenerative diseases.

These results suggest that a malfunction in the mechanisms regulatingtau O-GlcNAc levels may be vitally important in the formation of NFTsand associated neurodegeneration. Good support for blocking tauhyperphosphorylation as a therapeutically useful intervention⁶² comesfrom recent studies showing that when transgenic mice harbouring humantau are treated with kinase inhibitors, they do not develop typicalmotor defects³⁴ and, in another case,³³ show decreased levels ofinsoluble tau. These studies provide a clear link between lowering tauphosphorylation levels and alleviating AD-like behavioural symptoms in amurine model of this disease. Indeed, pharmacological modulation of tauhyperphosphorylation is widely recognized as a valid therapeuticstrategy for treating AD and other neurodegenerative disorders.⁶³

Small-molecule O-GlcNAcase inhibitors, to limit tauhyperphosphorylation, have been considered for treatment of AD andrelated tauopathies.⁶⁴ Specifically, the O-GlcNAcase inhibitor thiamet-Ghas been implicated in the reduction of tau phosphorylation in culturedPC-12 cells at pathologically relevant sites.⁶⁴ Moreover, oraladministration of thiamet-G to healthy Sprague-Dawley rats has beenimplicated in reduced phosphorylation of tau at Thr231, Ser396 andSer422 in both rat cortex and hippocampus.⁶⁴

There is also a large body of evidence indicating that increased levelsof O-GlcNAc protein modification provides protection against pathogeniceffects of stress in cardiac tissue, including stress caused byischemia, hemorrhage, hypervolemic shock, and calcium paradox. Forexample, activation of the hexosamine biosynthetic pathway (HBP) byadministration of glucosamine has been demonstrated to exert aprotective effect in animals models of ischemia/reperfusion,⁶⁵⁻⁷¹ traumahemorrhage,⁷²⁻⁷⁴ hypervolemic shock,⁷⁵ and calcium paradox.^(65,76)Moreover, strong evidence indicates that these cardioprotective effectsare mediated by elevated levels of protein O-GlcNAcmodification.^(65,66,68,71,73,76-79) There is also evidence that theO-GlcNAc modification plays a role in a variety of neurodegenerativediseases, including Parkinson's disease and Huntington's disease.⁸⁰

Humans have three genes encoding enzymes that cleave terminalβ-N-acetylglucosamine residues from glycoconjugates. The first of theseencodes O-GlcNAcase. O-GlcNAcase is a member of family 84 of glycosidehydrolases that includes enzymes from organisms as diverse asprokaryotic pathogens to humans (for the family classification ofglycoside hydrolases see Coutinho, P. M. & Henrissat, B. (1999)Carbohydrate-Active Enzymes server at URL:http://afmb.cnrs-mrs.fr/CAZY/.^(81,82) O-GlcNAcase acts to hydrolyseO-GlcNAc off of serine and threonine residues of post-translationallymodified proteins.^(1,6,7,83,84) Consistent with the presence ofO-GlcNAc on many intracellular proteins, the enzyme O-GlcNAcase appearsto have a role in the etiology of several diseases including type IIdiabetes,^(14,85) AD^(16,21,86) and cancer.^(22,87) Although O-GlcNAcasewas likely isolated earlier on,^(18,19) about 20 years elapsed beforeits biochemical role in acting to cleave O-GlcNAc from serine andthreonine residues of proteins was understood.⁶ More recentlyO-GlcNAcase has been cloned,⁷ partially characterized,²⁰ and suggestedto have additional activity as a histone acetyltransferase.²⁰ However,little was known about the catalytic mechanism of this enzyme.

The other two genes, HEXA and HEXB, encode enzymes catalyzing thehydrolytic cleavage of terminal β-N-acetylglucosamine residues fromglycoconjugates. The gene products of HEXA and HEXB predominantly yieldtwo dimeric isozymes, hexosaminidase A and hexosaminidase B,respectively. Hexosaminidase A (αβ), a heterodimeric isozyme, iscomposed of an α- and a β-subunit. Hexosaminidase B (ββ), a homodimericisozyme, is composed of two β-subunits. The two subunits, α- and β-,bear a high level of sequence identity. Both of these enzymes areclassified as members of family 20 of glycoside hydrolases and arenormally localized within lysosomes. The proper functioning of theselysosomal β-hexosaminidases is critical for human development, a factthat is underscored by the tragic genetic illnesses, Tay-Sach's andSandhoff diseases which stem from a dysfunction in, respectively,hexosaminidase A and hexosaminidase B.⁸⁸ These enzymatic deficienciescause an accumulation of glycolipids and glycoconjugates in thelysosomes resulting in neurological impairment and deformation. Thedeleterious effects of accumulation of gangliosides at the organismallevel are still being uncovered.⁸⁹

As a result of the biological importance of theseβ-N-acetyl-glucosaminidases, small molecule inhibitors ofglycosidases⁹⁰⁻⁹³ have received a great deal of attention,⁹⁴ both astools for elucidating the role of these enzymes in biological processesand in developing potential therapeutic applications. The control ofglycosidase function using small molecules offers several advantagesover genetic knockout studies including the ability to rapidly varydoses or to entirely withdraw treatment.

However, a major challenge in developing inhibitors for blocking thefunction of mammalian glycosidases, including O-GlcNAcase, is the largenumber of functionally related enzymes present in tissues of highereukaryotes. Accordingly, the use of non-selective inhibitors in studyingthe cellular and organismal physiological role of one particular enzymeis complicated because complex phenotypes arise from the concomitantinhibition of such functionally related enzymes. In the case ofβ-N-acetylglucosaminidases, many compounds that act to block O-GlcNAcasefunction are non-specific and act potently to inhibit the lysosomalβ-hexosaminidases.

A few of the better characterized inhibitors ofβ-N-acetyl-glucosaminidases which have been used in studies of O-GlcNAcpost-translational modification within both cells and tissues arestreptozotocin (STZ), 2′-methyl-α-D-glucopyrano-[2,1-d]-Δ2′-thiazoline(NAG-thiazoline) and O-(2-acetamido-2-deoxy-D-glucopyranosylidene)aminoN-phenylcarbamate (PUGNAc).^(14,95-98)

STZ has long been used as a diabetogenic compound because it has aparticularly detrimental effect on β-islet cells.⁹⁹ STZ exerts itscytotoxic effects through both the alkylation of cellular DNA^(99,100)as well as the generation of radical species including nitric oxide.¹⁰¹The resulting DNA strand breakage promotes the activation ofpoly(ADP-ribose) polymerase (PARP)¹⁰² with the net effect of depletingcellular NAD+ levels and, ultimately, leading to cell death.^(103,104)Other investigators have proposed instead that STZ toxicity is aconsequence of the irreversible inhibition of O-GlcNAcase, which ishighly expressed within β-islet cells.^(95,105) This hypothesis has,however, been brought into question by two independent researchgroups.^(106,107) Because cellular O-GlcNAc levels on proteins increasein response to many forms of cellular stress¹⁰⁸ it seems possible thatSTZ results in increased O-GlcNAc-modification levels on proteins byinducing cellular stress rather than through any specific and directaction on O-GlcNAcase. Indeed, Hanover and coworkers have shown that STZfunctions as a poor and somewhat selective inhibitor of O-GlcNAcase¹⁰⁹and although it has been proposed by others that STZ acts toirreversibly inhibit O-GlcNAcase,¹¹⁰ there has been no cleardemonstration of this mode of action. More recently, it has been shownthat STZ does not irreversibly inhibit O-GlcNAcase.¹¹¹

NAG-thiazoline has been found to be a potent inhibitor of family 20hexosaminidases,^(93,112) and more recently, the family 84O-GlcNAcases.¹¹¹ Despite its potency, a downside to using NAG-thiazolinein a complex biological context is that it lacks selectivity andtherefore perturbs multiple cellular processes.

PUGNAc is another compound that suffers from the same problem of lack ofselectivity, yet has enjoyed use as an inhibitor of both humanO-GlcNAcase^(6,113) and the family 20 human β-hexosaminidases.¹¹⁴ Thismolecule, developed by Vasella and coworkers, was found to be a potentcompetitive inhibitor of the β-N-acetyl-glucosaminidases from Canavaliaensiformis, Mucor rouxii, and the β-hexosaminidase from bovine kidney.⁹¹It has been demonstrated that administration of PUGNAc in a rat model oftrauma hemorrhage decreases circulating levels of the pro-inflammatorycytokines TNF-α and IL-6.¹¹⁵ It has also been shown that administrationof PUGNAc in a cell-based model of lymphocyte activation decreasesproduction of the cytokine IL-2.¹¹⁶ Subsequent studies have indicatedthat PUGNAc can be used in an animal model to reduce myocardial infarctsize after left coronary artery occlusions.¹¹⁷ Of particularsignificance is the fact that elevation of O-GlcNAc levels byadministration of PUGNAc, an inhibitor of O-GlcNAcase, in a rat model oftrauma hemorrhage improves cardiac function.^(115,118) In addition,elevation of O-GlcNAc levels by treatment with PUGNAc in a cellularmodel of ischemia/reperfusion injury using neonatal rat ventricularmyocytes improved cell viability and reduced necrosis and apoptosiscompared to untreated cells.¹¹⁹

More recently, it has been suggested that the selective O-GlcNAcaseinhibitor NButGT exhibits protective activity in cell-based models ofischemia/reperfusion and cellular stresses, including oxidativestress.¹²⁰ This study suggests the use of O-GlcNAcase inhibitors toelevate protein O-GlcNAc levels and thereby prevent the pathogeniceffects of stress in cardiac tissue.

International patent applications PCT/CA2006/000300, filed 1 Mar. 2006,published under No. WO 2006/092049 on 8 Sep. 2006; PCT/CA2007/001554,filed 31 Aug. 2007, published under No. WO 2008/025170 on 6 Mar. 2008;PCT/CA2009/001087, filed 31 Jul. 2009, published under No. WO2010/012106 on 4 Feb. 2010; PCT/CA2009/001088, filed 31 Jul. 2009,published under WO 2010/012107 on 4 Feb. 2010; PCT/CA2009/001302, filed16 Sep. 2009, published under WO 2010/037207 on 8 Apr. 2010;PCT/CA2011/000548, filed 10 May 2011, published under No. WO 2011/140640on 17 Nov. 2011; PCT/CA/2011/001241, filed 8 Nov. 2011, published underWO 2012/061927 on 18 May 2012; PCT/US2011/059668, filed 8 Nov. 2011,published under WO 2012/064680 on 18 May 2012; and PCT/CA2011/001397,filed 21 Dec. 2011, published under WO 2012/083435 on 28 Jun. 2012,describe selective inhibitors of O-GlcNAcase.

SUMMARY OF THE INVENTION

The invention provides, in part, compounds for inhibiting glycosidases,prodrugs of the compounds, uses of the compounds and the prodrugs,pharmaceutical compositions including the compounds or prodrugs of thecompounds, and methods of treating diseases and disorders related todeficiency or overexpression of O-GlcNAcase, and/or accumulation ordeficiency of O-GlcNAc.

In one aspect, the invention provides a compound of Formula (I) or apharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; R⁶ may beselected from the group consisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH;and R⁷ may be selected from the group consisting of: H, C₁₋₁₈ alkyl,C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R², R³, R⁴, or R⁵ may be OH; or R¹ may beCH₂OH; or R⁶ may be CH₂OH.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R², R³, R⁴, or R⁵ may be F; or R¹ may beCH₂F or CHF₂; or R⁶ may be CH₂F or CHF₂.

In alternative embodiments, the invention provides a compound of Formula(I) in which R¹ may be CH₃; or R² and R³ may be H; or R⁴ and R⁵ may beH; or R⁶ may be CH₃.

In alternative embodiments, the invention provides a compound of Formula(I) in which R¹ or R⁶ may be independently H.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R¹ or R⁶ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(I) in which R¹ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(I) in which R⁶ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(I) in which R¹ and R⁶ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R² or R³ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R⁴ or R⁵ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R², R³, R⁴, or R⁵ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R¹, R², R³, R⁴, R⁵, or R⁶ may be other thanH.

In alternative embodiments, the invention provides a compound of Formula(I) in which at least one of R¹ or R⁶ may be other than CH₃.

In alternative embodiments, the invention provides a compound of Formula(I) in which R⁷ may be other than H.

In alternative embodiments, the invention provides a compound of Formula(Ia) or a pharmaceutically acceptable salt thereof:

where R⁸ may be H or C₁₋₆ alkyl; R⁹ may be C₁₋₆ acyl; R² may be H and R³may be OH, or R² may be H and R³ may be H, or R² may be H and R³ may beF, or R² may be F and R³ may be H, or R² may be F and R³ may be F, or R²may be OH and R³ may be H; R⁴ may be H and R⁵ may be OH, or R⁴ may be Hand R⁵ may be H, or R⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵may be H, or R⁴ may be F and R⁵ may be F, or R⁴ may be OH and R⁵ may beH; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F, CHF₂,and CH₂OH; and R⁷ may be selected from the group consisting of: H, C₁₋₁₈alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl,C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ib) or a pharmaceutically acceptable salt thereof:

where each R¹⁰ may be independently H or C₁₋₆ alkyl; R² may be H and R³may be OH, or R² may be H and R³ may be H, or R² may be H and R³ may beF, or R² may be F and R³ may be H, or R² may be F and R³ may be F, or R²may be OH and R³ may be H; R⁴ may be H and R⁵ may be OH, or R⁴ may be Hand R⁵ may be H, or R⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵may be H, or R⁴ may be F and R⁵ may be F, or R⁴ may be OH and R⁵ may beH; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F, CHF₂,and CH₂OH; and R⁷ may be selected from the group consisting of: H, C₁₋₁₈alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl,C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ic) or a pharmaceutically acceptable salt thereof:

where each R¹⁰ may be independently H or C₁₋₆ alkyl; R² may be H and R³may be OH, or R² may be H and R³ may be H, or R² may be H and R³ may beF, or R² may be F and R³ may be H, or R² may be F and R³ may be F, or R²may be OH and R³ may be H; R⁴ may be H and R⁵ may be OH, or R⁴ may be Hand R⁵ may be H, or R⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵may be H, or R⁴ may be F and R⁵ may be F, or R⁴ may be OH and R⁵ may beH; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F, CHF₂,and CH₂OH; and R⁷ may be selected from the group consisting of: H, C₁₋₁₈alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl,C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Id) or a pharmaceutically acceptable salt thereof:

where R² may be H and R³ may be OH, or R² may be H and R³ may be H, orR² may be H and R³ may be F, or R² may be F and R³ may be H, or R² maybe F and R³ may be F, or R² may be OH and R³ may be H; R⁴ may be H andR⁵ may be OH, or R⁴ may be H and R⁵ may be H, or R⁴ may be H and R⁵ maybe F, or R⁴ may be F and R⁵ may be H, or R⁴ may be F and R⁵ may be F, orR⁴ may be OH and R⁵ may be H; R⁶ may be selected from the groupconsisting of H, CH₃, CH₂F, CHF₂, and CH₂OH; and R⁷ may be selected fromthe group consisting of H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl,C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ie) or a pharmaceutically acceptable salt thereof:

where R² may be H and R³ may be OH, or R² may be H and R³ may be H, orR² may be H and R³ may be F, or R² may be F and R³ may be H, or R² maybe F and R³ may be F, or R² may be OH and R³ may be H; R⁴ may be H andR⁵ may be OH, or R⁴ may be H and R⁵ may be H, or R⁴ may be H and R⁵ maybe F, or R⁴ may be F and R⁵ may be H, or R⁴ may be F and R⁵ may be F, orR⁴ may be OH and R⁵ may be H; R⁶ may be selected from the groupconsisting of H, CH₃, CH₂F, CHF₂, and CH₂OH; and R⁷ may be selected fromthe group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl,C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(If) or a pharmaceutically acceptable salt thereof:

where R² may be H and R³ may be OH, or R² may be H and R³ may be H, orR² may be H and R³ may be F, or R² may be F and R³ may be H, or R² maybe F and R³ may be F, or R² may be OH and R³ may be H; R⁴ may be H andR⁵ may be OH, or R⁴ may be H and R⁵ may be H, or R⁴ may be H and R⁵ maybe F, or R⁴ may be F and R⁵ may be H, or R⁴ may be F and R⁵ may be F, orR⁴ may be OH and R⁵ may be H; R⁶ may be selected from the groupconsisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH; and R⁷ may be selectedfrom the group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl,C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl,each excluding H optionally substituted from one to four substituentswith one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂,CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy,or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ig) or a pharmaceutically acceptable salt thereof:

where R² may be H and R³ may be OH, or R² may be H and R³ may be H, orR² may be H and R³ may be F, or R² may be F and R³ may be H, or R² maybe F and R³ may be F, or R² may be OH and R³ may be H; R⁴ may be H andR⁵ may be OH, or R⁴ may be H and R⁵ may be H, or R⁴ may be H and R⁵ maybe F, or R⁴ may be F and R⁵ may be H, or R⁴ may be F and R⁵ may be F, orR⁴ may be OH and R⁵ may be H; R⁶ may be selected from the groupconsisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH; and R⁷ may be selectedfrom the group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl,C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl,each excluding H optionally substituted from one to four substituentswith one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂,CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy,or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ih) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R⁴ may be H and R⁵ may be OH, or R⁴ maybe H and R⁵ may be H, or R⁴ may be H and R⁵ may be F, or R⁴ may be F andR⁵ may be H, or R⁴ may be F and R⁵ may be F, or R⁴ may be OH and R⁵ maybe H; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F,CHF₂, and CH₂OH; and R⁷ may be selected from the group consisting of: H,C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ii) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R⁴ may be H and R⁵ may be OH, or R⁴ maybe H and R⁵ may be H, or R⁴ may be H and R⁵ may be F, or R⁴ may be F andR⁵ may be H, or R⁴ may be F and R⁵ may be F, or R⁴ may be OH and R⁵ maybe H; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F,CHF₂, and CH₂OH; and R⁷ may be selected from the group consisting of: H,C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ij) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R⁴ may be H and R⁵ may be OH, or R⁴ maybe H and R⁵ may be H, or R⁴ may be H and R⁵ may be F, or R⁴ may be F andR⁵ may be H, or R⁴ may be F and R⁵ may be F, or R⁴ may be OH and R⁵ maybe H; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F,CHF₂, and CH₂OH; and R⁷ may be selected from the group consisting of: H,C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ik) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F,CHF₂, and CH₂OH; and R⁷ may be selected from the group consisting of: H,C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Il) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F,CHF₂, and CH₂OH; and R⁷ may be selected from the group consisting of: H,C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Im) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F,CHF₂, and CH₂OH; and R⁷ may be selected from the group consisting of: H,C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃,CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(In) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; and R⁷ may beselected from the group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Io) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; and R⁷ may beselected from the group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Ip) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; and R⁷ may beselected from the group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(Iq) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; and R⁷ may beselected from the group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In alternative embodiments, the invention provides a compound of Formula(II) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; R⁶ may beselected from the group consisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH;and R⁷ may be selected from the group consisting of: C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each optionally substituted from one to foursubstituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl; with the proviso that R⁷excludes benzyl, 4-hydroxybenzyl, 4-(pyridin-2-yl)benzyl, (CH₂)₂phenyl,C(O)CH₃, [1,1′-biphenyl]-4-ylmethyl, (R)-1-phenylethyl,(S)-1-phenylethyl, CH(phenyl)₂, and 4-fluorophenethyl.

In alternative embodiments, the compound may be a prodrug; the compoundmay selectively inhibit an O-glycoprotein2-acetamido-2-deoxy-β-D-glucopyranosidase (0-GlcNAcase); the compoundmay selectively bind an O-GlcNAcase (e.g., a mammalian O-GlcNAcase); thecompound may selectively inhibit the cleavage of a2-acetamido-2-deoxy-β-D-glucopyranoside (O-GlcNAc); the compound may notsubstantially inhibit a mammalian β-hexosaminidase.

In alternative embodiments, a compound according to Formula (I), Formula(Ia), Formula (Ib), Formula (Ic), Formula (Id), Formula (Ie), Formula(If), Formula (Ig), Formula (Ih), Formula (Ii), Formula (Ij), Formula(Ik), Formula (Il), Formula (Im), Formula (In), Formula (Io), Formula(Ip), or Formula (Iq) may have enhanced permeability.

In alternative embodiments, a compound according to Formula (Ia),Formula (Ib), or Formula (Ic) may have enhanced selectivity.

In alternative embodiments, a compound according to Formula (If),Formula (Ig), Formula (Ih), Formula (Ii), Formula (Ik), Formula (Il),Formula (In), or Formula (Io) may have enhanced permeability.

In alternative embodiments, a compound according to Formula (Id),Formula (Ie), Formula (Ij), Formula (Im), Formula (Ip), or Formula (Iq)may have enhanced permeability.

In alternative aspects, the invention provides a pharmaceuticalcomposition including a compound according to the invention, incombination with a pharmaceutically acceptable carrier.

In alternative aspects, the invention provides methods of selectivelyinhibiting an O-GlcNAcase, or of inhibiting an O-GlcNAcase in a subjectin need thereof, or of increasing the level of O-GlcNAc, or of treatinga neurodegenerative disease, a tauopathy, cancer or stress, in a subjectin need thereof, by administering to the subject an effective amount ofa compound of Formula (I) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; R⁶ may beselected from the group consisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH;and R⁷ may be selected from the group consisting of: H, C₁₋₁₈ alkyl,C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl. The condition may be Alzheimer'sdisease, Amyotrophic lateral sclerosis (ALS), Amyotrophic lateralsclerosis with cognitive impairment (ALSci), Argyrophilic graindementia, Bluit disease, Corticobasal degeneration (CBD), Dementiapugilistica, Diffuse neurofibrillary tangles with calcification, Down'ssyndrome, Familial British dementia, Familial Danish dementia,Frontotemporal dementia with parkinsonism linked to chromosome 17(FTDP-17), Gerstmann-Straussler-Scheinker disease, Guadeloupeanparkinsonism, Hallevorden-Spatz disease (neurodegeneration with brainiron accumulation type 1), Multiple system atrophy, Myotonic dystrophy,Niemann-Pick disease (type C), Pallido-ponto-nigral degeneration,Parkinsonism-dementia complex of Guam, Pick's disease (PiD),Post-encephalitic parkinsonism (PEP), Prion diseases (includingCreutzfeldt-Jakob Disease (CJD), Variant Creutzfeldt-Jakob Disease(vCJD), Fatal Familial Insomnia, and Kuru), Progressive supercorticalgliosis, Progressive supranuclear palsy (PSP), Richardson's syndrome,Subacute sclerosing panencephalitis, Tangle-only dementia, Huntington'sdisease, Parkinson's disease, Schizophrenia, Mild Cognitive Impairment(MCI), Neuropathy (including peripheral neuropathy, autonomicneuropathy, neuritis, and diabetic neuropathy), or Glaucoma. The stressmay be a cardiac disorder, e.g., ischemia; hemorrhage; hypovolemicshock; myocardial infarction; an interventional cardiology procedure;cardiac bypass surgery; fibrinolytic therapy; angioplasty; or stentplacement.

In alternative aspects, the invention provides a method of treating anO-GlcNAcase-mediated condition that excludes a neurodegenerativedisease, a tauopathy, cancer or stress, in a subject in need thereof, byadministering to the subject an effective amount of a compound ofFormula (I) or a pharmaceutically acceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; R⁶ may beselected from the group consisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH;and R⁷ may be selected from the group consisting of: H, C₁₋₁₈ alkyl,C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl. In some embodiments, thecondition may be inflammatory or allergic diseases such as asthma,allergic rhinitis, hypersensitivity lung diseases, hypersensitivitypneumonitis, eosinophilic pneumonias, delayed-type hypersensitivity,atherosclerosis, interstitial lung disease (ILD) (e.g., idiopathicpulmonary fibrosis, or ILD associated with rheumatoid arthritis,systemic lupus erythematosus, ankylosing spondylitis, systemicsclerosis, Sjogren's syndrome, polymyositis or dermatomyositis);systemic anaphylaxis or hypersensitivity responses, drug allergies,insect sting allergies; autoimmune diseases, such as rheumatoidarthritis, psoriatic arthritis, multiple sclerosis, Guillain-Barrésyndrome, systemic lupus erythematosus, myastenia gravis,glomerulonephritis, autoimmune thyroiditis, graft rejection, includingallograft rejection or graft-versus-host disease; inflammatory boweldiseases, such as Crohn's disease and ulcerative colitis;spondyloarthropathies; scleroderma; psoriasis (including T-cell mediatedpsoriasis) and inflammatory dermatoses such as dermatitis, eczema,atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis(e.g., necrotizing, cutaneous, and hypersensitivity vasculitis);eosinphilic myotis, and eosiniphilic fasciitis; graft rejection, inparticular but not limited to solid organ transplants, such as heart,lung, liver, kidney, and pancreas transplants (e.g. kidney and lungallografts); epilepsy; pain; fibromyalgia; stroke, e.g., neuroprotectionfollowing a stroke.

In alternative embodiments, the administering may increase the level ofO-GlcNAc in the subject. The subject may be a human.

In alternative aspects, the invention provides use of a compound of aneffective amount of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof:

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; R⁶ may beselected from the group consisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH;and R⁷ may be selected from the group consisting of: H, C₁₋₁₈ alkyl,C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl, in the preparation of amedicament. The medicament may be for selectively inhibiting anO-GlcNAcase, for increasing the level of O-GlcNAc, for treating acondition modulated by an O-GlcNAcase, for treating a neurodegenerativedisease, a tauopathy, a cancer, or stress.

In alternative aspects, the invention provides a method for screeningfor a selective inhibitor of an O-GlcNAcase, by a) contacting a firstsample with a test compound; b) contacting a second sample with acompound of Formula (I)

where R¹ may be selected from the group consisting of: CH₂NR⁸R⁹,(CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, and CH₂OH, where R⁸ may be H orC₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0 or 1, and each R¹⁰ may beindependently H or C₁₋₆ alkyl; R² may be H and R³ may be OH, or R² maybe H and R³ may be H, or R² may be H and R³ may be F, or R² may be F andR³ may be H, or R² may be F and R³ may be F, or R² may be OH and R³ maybe H; R⁴ may be H and R⁵ may be OH, or R⁴ may be H and R⁵ may be H, orR⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵ may be H, or R⁴ maybe F and R⁵ may be F, or R⁴ may be OH and R⁵ may be H; R⁶ may beselected from the group consisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH;and R⁷ may be selected from the group consisting of: H, C₁₋₁₈ alkyl,C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl; c) determining the level ofinhibition of the O-GlcNAcase in the first and second samples, where thetest compound is a selective inhibitor of a O-GlcNAcase if the testcompound exhibits the same or greater inhibition of the O-GlcNAcase whencompared to the compound of Formula (I).

This summary of the invention does not necessarily describe all featuresof the invention.

DETAILED DESCRIPTION

The invention provides, in part, novel compounds that are capable ofinhibiting an O-glycoprotein 2-acetamido-2-deoxy-β-D-glucopyranosidase(O-GlcNAcase). In some embodiments, the O-GlcNAcase may be a mammalianO-GlcNAcase, such as a rat, mouse or human O-GlcNAcase.

In some embodiments, one or more of the compounds according to theinvention may exhibit enhanced permeability. Permeability can beassessed using a variety of standard experimental techniques, includingwithout limitation in situ perfusion, ex vivo tissue diffusion, in vitrocell monolayers (e.g. Caco-2 cells, MDCK cells, LLC-PK1 cells), andartificial cell membranes (e.g. PAMPA assay); suitable techniques formeasuring effective permeability (P_(eff)) or apparent peameability(P_(app)) are reviewed for example by Volpe in The AAPSJournal, 2010,12(4), 670-678. In some embodiments, one or more of the compoundsaccording to the invention may show enhanced permeability when tested inone or more of these assays for determining P_(eff) or P_(app). In someembodiments, a compound that exhibits enhanced permeability may exhibitgreater oral absorption. In some embodiments, a compound that exhibitsenhanced permeability may exhibit greater brain penetrance whenadministered in vivo. In some embodiments, a compound that exhibitsenhanced permeability may achieve higher brain concentrations whenadministered in vivo. In some embodiments, a compound that exhibitsenhanced permeability may exhibit a higher brain/plasma concentrationratio when administered in vivo. In some embodiments, “enhancedpermeability” means an increase in measured P_(eff) or P_(app) by anyvalue between 10% and 100%, or of any integer value between 10% and100%, for example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, orover 100%, or an increase by 1-fold, 2-fold, or 3-fold, or more, ascompared to a suitable reference compound disclosed in for example WO2006/092049 or WO 2008/025170. A suitable reference compound may be, forexample,(3aR,5R,6S,7R,7aR)-5-(hydroxymethyl)-2-propyl-5,6,7,7a-tetrahydro-3aH-pyrano[3,2-d]thiazole-6,7-diol,or(3aR,5R,6S,7R,7aR)-2-(ethylamino)-5-(hydroxymethyl)-5,6,7,7a-tetrahydro-3aH-pyrano[3,2-d]thiazole-6,7-diol,or(3aR,5R,6S,7R,7aR)-2-(dimethylamino)-5-(hydroxymethyl)-5,6,7,7a-tetrahydro-3aH-pyrano[3,2-d]thiazole-6,7-diol.In some embodiments, “enhanced permeability” means a measurable P_(app)value (i.e. a value greater than zero) in the assay described below fordetermination of P_(app) in LLC-PK1 cells. In some embodiments,“enhanced permeability” means a P_(app) value greater than 2×10⁻⁶ cm/sin the assay described below for determination of P_(app) in LLC-PK1cells. In alternative embodiments, “enhanced permeability” means aP_(app) value in the range 2×10⁻⁶ cm/s to 35×10⁻⁶ cm/s in the assaydescribed below for determination of P_(app) in LLC-PK1 cells.

In some embodiments, a compound according to the invention may exhibitsuperior selectivity in inhibiting an O-GlcNAcase. In some embodiments,one or more of the compounds according to the invention may be moreselective for an O-GlcNAcase over a β-hexosaminidase. In someembodiments, one or more of the compounds may selectively inhibit theactivity of a mammalian O-GlcNAcase over a mammalian β-hexosaminidase.In some embodiments, a selective inhibitor of an O-GlcNAcase may notsubstantially inhibit a β-hexosaminidase. In some embodiments, theβ-hexosaminidase may be a mammalian β-hexosaminidase, such as a rat,mouse or human β-hexosaminidase. A compound that “selectively” inhibitsan O-GlcNAcase is a compound that may inhibit the activity or biologicalfunction of an O-GlcNAcase, but may not substantially inhibit theactivity or biological function of a β-hexosaminidase. For example, insome embodiments, a selective inhibitor of an O-GlcNAcase mayselectively inhibit the cleavage of2-acetamido-2-deoxy-β-D-glucopyranoside (O-GlcNAc) from polypeptides. Insome embodiments, a selective inhibitor of an O-GlcNAcase mayselectively bind to an O-GlcNAcase. In some embodiments, a selectiveinhibitor of an O-GlcNAcase may inhibit hyperphosphorylation of a tauprotein and/or inhibit formations of NFTs. By “inhibit,” “inhibition” or“inhibiting” means a decrease by any value between 10% and 90%, or ofany integer value between 30% and 60%, or over 100%, or a decrease by1-fold, 2-fold, 5-fold, 10-fold or more. It is to be understood that theinhibiting does not require full inhibition. In some embodiments, aselective inhibitor of an O-GlcNAcase may elevate or enhance O-GlcNAclevels e.g., O-GlcNAc-modified polypeptide or protein levels, in cells,tissues, or organs (e.g., in brain, muscle, or heart (cardiac) tissue)and in animals. By “elevating” or “enhancing” is meant an increase byany value between 10% and 90%, or of any integer value between 30% and60%, or over 100%, or an increase by 1-fold, 2-fold, 5-fold, 10-fold,15-fold, 25-fold, 50-fold, 100-fold or more. In some embodiments, aselective inhibitor of an O-GlcNAcase may exhibit a selectivity ratio,as described herein, in the range 10 to 100000, or in the range 100 to100000, or in the range 1000 to 100000, or at least 10, 20, 50, 100,200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 6000,7000, 10,000, 25,000, 50,000, 75,000, or any value within or about thedescribed range.

One or more of the compounds of the present invention may elevateO-GlcNAc levels on O-GlcNAc-modified polypeptides or proteins in vivospecifically via interaction with an O-GlcNAcase enzyme, and may beeffective in treating conditions which require or respond to inhibitionof O-GlcNAcase activity.

In some embodiments, one or more of the compounds of the presentinvention may be useful as agents that produce a decrease in tauphosphorylation and NFT formation. In some embodiments, one or more ofthe compounds may therefore be useful to treat Alzheimer's disease andrelated tauopathies. In some embodiments, one or more of the compoundsmay thus be capable of treating Alzheimer's disease and relatedtauopathies by lowering tau phosphorylation and reducing NFT formationas a result of increasing tau O-GlcNAc levels. In some embodiments, oneor more of the compounds may produce an increase in levels of O-GlcNAcmodification on O-GlcNAc-modified polypeptides or proteins, and maytherefore be useful for treatment of disorders responsive to suchincreases in O-GlcNAc modification; these disorders may include, withoutlimitation, neurodegenerative, inflammatory, cardiovascular, andimmunoregulatory diseases. In some embodiments, a compound may also beuseful as a result of other biological activities related to its abilityto inhibit the activity of glycosidase enzymes. In alternativeembodiments, one or more of the compounds of the invention may bevaluable tools in studying the physiological role of O-GlcNAc at thecellular and organismal level.

In alternative embodiments, the invention provides methods of enhancingor elevating levels of protein O-GlcNAc modification in animal subjects,such as, veterinary and human subjects. In alternative embodiments, theinvention provides methods of selectively inhibiting an O-GlcNAcaseenzyme in animal subjects, such as, veterinary and human subjects. Inalternative embodiments, the invention provides methods of inhibitingphosphorylation of tau polypeptides, or inhibiting formation of NFTs, inanimal subjects, such as, veterinary and human subjects.

In specific embodiments, the invention provides compounds describedgenerally by Formula (I) and the salts, prodrugs, and enantiomeric formsthereof:

As set forth in Formula (I): R¹ may be selected from the groupconsisting of: CH₂NR⁸R⁹, (CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F, CHF₂, andCH₂OH, where R⁸ may be H or C₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, n may be 0or 1, and each R¹⁰ may be independently H or C₁₋₆ alkyl; R² may be H andR³ may be OH, or R² may be H and R³ may be H, or R² may be H and R³ maybe F, or R² may be F and R³ may be H, or R² may be F and R³ may be F, orR² may be OH and R³ may be H; R⁴ may be H and R⁵ may be OH, or R⁴ may beH and R⁵ may be H, or R⁴ may be H and R⁵ may be F, or R⁴ may be F and R⁵may be H, or R⁴ may be F and R⁵ may be F, or R⁴ may be OH and R⁵ may beH; R⁶ may be selected from the group consisting of: H, CH₃, CH₂F, CHF₂,and CH₂OH; and R⁷ may be selected from the group consisting of: H, C₁₋₁₈alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl,C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀ arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀arylalkynyl, C₂₋₂₀ heteroarylalkyl, C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀heteroarylalkynyl, each excluding H optionally substituted from one tofour substituents with one of more of halo, OH, OCH₃, OCF₃, CN, SO₂NH₂,SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl.

In some embodiments, R¹ as set forth in Formula (I) may be selected fromthe group consisting of: CH₂NR⁸R⁹, (CH₂)_(n)C(O)NR¹⁰ ₂, H, CH₃, CH₂F,CHF₂, and CH₂OH, where R⁸ may be H or C₁₋₆ alkyl, R⁹ may be C₁₋₆ acyl, nmay be 0 or 1, and each R¹⁰ may be independently H or C₁₋₆ alkyl. Insome embodiments, R¹ may be CH₂NR⁸R⁹, where R⁸ may be H or C₁₋₆ alkyl,and R⁹ may be C₁₋₆ acyl. In some embodiments, R¹ may be CH₂NR⁸R⁹, whereR⁸ may be H, and R⁹ may be C(O)CH₃, C(O)CH₂CH₃, or C(O)(CH₂)₂CH₃. Insome embodiments, R¹ may be C(O)NR¹⁰ ₂, where each R¹⁰ may beindependently H or C₁₋₆ alkyl. In some embodiments, R¹ may be(CH₂)C(O)NR¹⁰ ₂, where each R¹⁰ may be independently H or C₁₋₆ alkyl. Insome embodiments, R¹ may be C(O)NHR¹⁰, where R¹⁰ may be H or C₁₋₆ alkyl.In some embodiments, R¹ may be (CH₂)C(O)NHR¹⁰, where R¹⁰ may be H orC₁₋₆ alkyl. In some embodiments, R¹ may be selected from the groupconsisting of: CH₂NH(C(O)CH₃), CH₂NH(C(O)CH₂CH₃), C(O)NHCH₃,C(O)NHCH₂CH₃, C(O)NH(CH₂)₂CH₃, (CH₂)C(O)NHCH₃, and (CH₂)C(O)NHCH₂CH₃. Insome embodiments, R¹ may be CH₂OH. In some embodiments, R¹ may be CH₂F.In some embodiments, R¹ may be CHF₂. In some embodiments, R¹ may be CH₃.In some embodiments, R¹ may be H.

In some embodiments, R² as set forth in Formula (I) may be H, F, or OH.In some embodiments, R² may be H or F. In some embodiments, R² may be F.In some embodiments, R² may be OH.

In some embodiments, R³ as set forth in Formula (I) may be H, F, or OH.In some embodiments, R³ may be H or F. In some embodiments, R³ may be F.In some embodiments, R³ may be H.

In some embodiments, R⁴ as set forth in Formula (I) may be H, F, or OH.In some embodiments, R⁴ may be H or F. In some embodiments, R⁴ may be F.In some embodiments, R⁴ may be H.

In some embodiments, R⁵ as set forth in Formula (I) may be H, F, or OH.In some embodiments, R⁵ may be H or F. In some embodiments, R⁵ may be F.In some embodiments, R⁵ may be OH.

In some embodiments, R⁶ as set forth in Formula (I) may be selected fromthe group consisting of: H, CH₃, CH₂F, CHF₂, and CH₂OH. In someembodiments, R⁶ may be CH₂OH. In some embodiments, R⁶ may be CH₂F orCHF₂. In some embodiments, R⁶ may be CH₃. In some embodiments, R⁶ may beH.

In some embodiments, R⁷ as set forth in Formula (I) may be selected fromthe group consisting of: H, C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,C₁₋₁₀ acyl, C₈₋₂₀ arylalkylacyl, C₃₋₂₀ heteroarylalkylacyl, C₇₋₂₀arylalkyl, C₈₋₂₀ arylalkenyl, C₈₋₂₀ arylalkynyl, C₂₋₂₀ heteroarylalkyl,C₃₋₂₀ heteroarylalkenyl, and C₃₋₂₀ heteroarylalkynyl, each excluding Hoptionally substituted from one to four substituents with one of more ofhalo, OH, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F,CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl. In someembodiments, R⁷ may be selected from the group consisting of: C₁₋₁₈alkyl, C₇₋₂₀ arylalkyl, and C₂₋₂₀ heteroarylalkyl, each optionallysubstituted from one to four substituents with one of more of halo, OH,OCH₃, OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F,CH₂CF₃, CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl. In someembodiments, R⁷ may be selected from the group consisting of: H, CH₃,(CH₂)₂(phenyl), (CH₂)₃(phenyl), (CH₂)₄(phenyl),3-(4-methoxyphenyl)propyl, 3-([1,1′-biphenyl]-4-yl)propyl,2-([1,1′-biphenyl]-4-yl)ethyl, (CH₂)(3-pyridyl), (CH₂)(2-pyridyl),([1,1-biphenyl]-4-yl)methyl(CH₂)₂CH(phenyl)₂,3-(4′-sulfamoyl-[1,1-biphenyl]-4-yl)propyl, 3-phenylpropanoyl,3-(6-fluoropyridin-3-yl)propyl, 4,4,4-trifluorobutyl,3-(5-(trifluoromethoxy)benzo[d]thiazol-2-yl)propyl,(Z)-5,5,5-trifluoropent-3-en-1-yl, pent-4-yn-1-yl, butyryl,3-(6-fluoropyridin-3-yl)propanoyl, cinnamyl, 3-phenylprop-2-yn-1-yl,(E)-3-(6-fluoropyridin-3-yl)allyl, and3-(6-fluoropyridin-3-yl)prop-2-yn-1-yl.

In specific embodiments of the invention, compounds according to Formula(I) include the compounds described in Table 1.

TABLE 1 Example Name Structure 1 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3- phenylpropyl)pyrrolidin-2- yl)methyl)acetamide

2 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-(4-methoxyphenyl)propyl)pyrrolidin-2- yl)methyl)acetamide

3 N-(((2R,3R,4R,5R)-1-(3-([1,1′- biphenyl]-4-yl)propyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

4 N-(((2R,3R,4R,5R)-1-(2-([1,1′- biphenyl]-4-yl)ethyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

5 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(4-phenylbutyppyrrolidin-2- yl)methyl)acetamide

6 N-(((2R,3R,4R,5R)-1-(3,3- diphenylpropyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

7 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(pyridin-3-ylmethyl)pyrrolidin-2- yl)methyl)acetamide

8 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(pyridin-2-ylmethyl)pyrrolidin-2- yl)methyl)acetamide

9 N-(((2R,3R,4R,5R)-1-([1,1′- biphenyl]-4-ylmethyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

10 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

11 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-(4-methoxyphenyl)propyl)pyrrolidin-2- yl)-N-ethylacetamide

12 2-((2R,3R,4R,5R)-1-(3,3- diphenylpropyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- ethylacetamide

13 2-((2R,3S,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

14 2-((2R,3R,4R,5R)-1-(3-([1,1′- biphenyl]-4-yl)propyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- ethylacetamide

15 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-(4′-sulfamoyl-[1,1′-biphenyl]-4-yl)propyl)pyrrolidin- 2-yl)-N-methylacetamide

16 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-phenethylpyrrolidin-2-yl)-N- ethylacetamide

17 (2S,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-phenethyl-N-propylpyrrolidine-2-carboxamide

18 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-phenethylpyrrolidin-2- yl)methyl)propionamide

19 N-ethyl-2-((2R,3R,4R,5S)-5- (fluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin-2-yl)acetamide

20 N-ethyl-2-((2R,3R,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-phenethylpyrrolidin-2-yl)acetamide

21 N-ethyl-2-((2R,3S,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-phenethylpyrrolidin-2-yl)acetamide

22 2-((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin- 2-yl)-N-ethylacetamide

23 2-((2R,3R,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

24 2-((2R,3R,4R,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

25 2-((2R,3S,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

26 2-((2R,3R,4S,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

27 2-((2R,3R,4R,5S)-5-(fluoromethyl)- 3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

28 2-((2R,3R,4R,5S)-5-(difluoromethyl)- 3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

29 2-((2R,3R,4R,5R)-3,4-dihydroxy-5-methyl-1-(3-phenylpropyl)pyrrolidin- 2-yl)-N-methylacetamide

30 2-((2S,4S,5R)-4-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

31 2-((2R,3S,5S)-3-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

32 2-((2R,3R,4R)-3,4-dihydroxy-1-(3- phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide

33 2-((2R,3R,4R,5S)-3-fluoro-5- (fluoromethyl)-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

34 2-((2R,3S,4R,5S)-5-(difluoromethyl)- 3-fluoro-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

35 N-(((2R,3R,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

36 N-(((2R,3R,4R,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

37 N-(((2R,3S,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

38 N-(((2R,3R,4S,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

39 N-(((2R,3R,4R,5S)-5-(fluoromethyl)- 3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

40 N-(((2R,3R,4R,5S)-5- (difluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

41 N-(((2S,4S,5R)-4-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

42 N-(((2R,3S,5S)-3-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

43 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-methyl-1-(3-phenylpropyl)pyrrolidin- 2-yl)methyl)propionamide

44 N-(((2R,3R,4R)-3,4-dihydroxy-1-(3- phenylpropyl)pyrrolidin-2-yl)methyl)propionamide

45 N-(((2R,3R,4R,5S)-3-fluoro-5- (fluoromethyl)-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

46 N-(((2R,3S,4R,5S)-5- (difluoromethyl)-3-fluoro-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

47 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-phenylpropanoyl)pyrrolidin-2- yl)methyl)propionamide

48 N-(((2R,3R,4R,5R)-1-(3-(6- fluoropyridin-3-yl)propyl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2- yl)methyl)propionamide

49 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-2- yl)methyl)propionamide

50 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-(5-(trifluoromethoxy)benzo[d]thiazol-2- yl)propyl)pyrrolidin-2-yl)methyl)propionamide

51 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-((Z)-5,5,5-trifluoropent-3-en-1-yl)pyrrolidin-2- yl)-N-methylacetamide

52 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(pent-4-yn-1-yl)pyrrolidin-2-yl)-N- methylacetamide

53 2-((2R,3R,4R,5R)-1-butyryl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

54 2-((2R,3R,4R,5R)-1-(3-(6- fluoropyridin-3-yl)propanoyl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

55 2-((2R,3R,4R,5R)-1-cinnamyl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

56 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-phenylprop-2-yn-1-yl)pyrrolidin-2-yl)-N- methylacetamide

57 2-((2R,3R,4R,5R)-1-((E)-3-(6- fluoropyridin-3-yl)allyl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

58 2-((2R,3R,4R,5R)-1-(3-(6- fluoropyridin-3-yl)prop-2-yn-1-yl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

In alternative embodiments of the invention, compounds according toFormula (I) include one or more of the compounds described in Table 2.

In alternative embodiments of the invention, one or more of thecompounds described in Table 2 are specifically excluded from thecompounds described in Formula (I).

TABLE 2 Name Structure N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)propionamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)butyramide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)pentanamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)hexanamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-methylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-butyl-3,4- dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-hexyl-3,4- dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-heptyl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-octylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-nonylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3S,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-nonylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- methylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)isobutyramide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)-3-methylbutanamide

N-(((2R,3R,4R,5R)-1-ethyl-3,4- dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-propylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-isopropylpyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-benzyl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-1- (4-hydroxybenzyl)-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(4-(pyridin-2-yl)benzyl)pyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-phenethylpyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-methylpyrrolidin-2-yl)methyl)propionamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)-N-methylacetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-methylpyrrolidin-2-yl)methyl)-N-methylacetamide

N-(((2R,3R,4R,5R)-1-acetyl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4S,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3S,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- methylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4S)-3,4- dihydroxypyrrolidin-2- yl)methyl)acetamide

N-(((2R,3S,4S)-3,4- dihydroxypyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4S)-3,4-dihydroxy-1- methylpyrrolidin-2- yl)methyl)acetamide

N-(((2R,3S,4S)-3,4- dihydroxypyrrolidin-2- yl)methyl)pivalamide

N-(((2R,3R,4S)-3,4-dihydroxy-1-(2- hydroxyethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3S,4S)-3,4- dihydroxypyrrolidin-2- yl)methyl)hexanamide

N-(((2R,3R,4S)-1-butyl-3,4- dihydroxypyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4S)-1-butyl-3,4- dihydroxypyrrolidin-2- yl)methyl)butyramide

N-(((2R,3R,4S)-1-benzyl-3,4- dihydroxypyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4S)-3,4-dihydroxy-1- nonylpyrrolidin-2- yl)methyl)acetamide

N-(((2R,3R,4S)-3,4-dihydroxy-1-(9- hydroxynonyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4S)-1-benzyl-3,4- dihydroxypyrrolidin-2- yl)methyl)butyramide

N-(((2R,3R,4S)-1-([1,1′-biphenyl]-4-ylmethyl)-3,4-dihydroxypyrrolidin-2- yl)methyl)acetamide

(2S,3S,4S)-3,4-dihydroxypyrrolidine- 2-carboxamide

(2S,3R,4R)-3,4-dihydroxypyrrolidine- 2-carboxamide

(2S)-3,4-dihydroxy-N,N- dimethylpyrrolidine-2-carboxamide

(2S,3R,4R)-N-(tert-butyl)-3,4- dihydroxypyrrolidine-2-carboxamide

N-butyl-2-((2R,3S,4R,5R)-3,4- dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)acetamide

N-butyl-2-((2R,3S,4R,5R)-3,4- dihydroxy-1-(2-hydroxyethyl)-5-(hydroxymethyl)pyrrolidin-2- yl)acetamide

N-butyl-2-((2R,3S,4R,5R)-1-butyl- 3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2- yl)acetamide

N-butyl-2-((2R,3S,4R,5R)-3,4- dihydroxy-5-(hydroxymethyl)-1-(9-hydroxynonyl)pyrrolidin-2- yl)acetamide

2-((2R,3S,4R,5R)-1-benzyl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- butylacetamide

2-((2R,3S,4R,5R)-1-([1,1′-biphenyl]- 4-ylmethyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- butylacetamide

(2S,3R,4R,5R)-2-(fluoromethyl)-5- (hydroxymethyl)pyrrolidine-3,4-diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3S,4R,5R)-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3S,4S,5R)-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1- methylpyrrolidine-3,4-diol

(2R,3R,4S,5R)-2,5- bis(hydroxymethyl)-1- methylpyrrolidine-3,4-diol

(2R,3R,4R,5R)-1-(2-hydroxyethyl)- 2,5-bis(hydroxymethyl)pyrrolidine-3,4-diol

(2R,3S,4R,5R)-1-(2-hydroxyethyl)- 2,5-bis(hydroxymethyl)pyrrolidine-3,4-diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1- propylpyrrolidine-3,4-diol

(2R,3R,4R,5R)-1-butyl-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1-(3-hydroxypropyl)pyrrolidine-3,4-diol

(2R,3S,4S,5R)-1-hexyl-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3R,4R,5R)-1-benzyl-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3S,4S,5R)-2,5- bis(hydroxymethyl)-1- octylpyrrolidine-3,4-diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1- nonylpyrrolidine-3,4-diol

(2R,3S,4S,5R)-1-decyl-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1-(9-hydroxynonyl)pyrrolidine-3,4-diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1-((R)-1-phenylethyl)pyrrolidine-3,4-diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1-((S)-1-phenylethyl)pyrrolidine-3,4-diol

(2R,3S,4S,5R)-1-hexadecyl-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3R,4R,5R)-2,5- bis(hydroxymethyl)-1- octadecylpyrrolidine-3,4-diol

(2R,3R,4R,5R)-1-benzhydryl-2,5- bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3R,4R,5R)-1-([1,1′-biphenyl]-4- ylmethyl)-2,5-bis(hydroxymethyl)pyrrolidine-3,4- diol

(2R,3R,4R,5R)-1-(4-fluorophenethyl)- 2,5-bis(hydroxymethyl)pyrrolidine-3,4-diol

(2R,3R,4R,5R)-2-(hydroxymethyl)-5- methylpyrrolidine-3,4-diol

(2R,3S,4R,5R)-2-(hydroxymethyl)-5- methylpyrrolidine-3,4-diol

(2R,3R,4S,5R)-2-(hydroxymethyl)-5- methylpyrrolidine-3,4-diol

(2R,3S,4S,5R)-2-(hydroxymethyl)-5- methylpyrrolidine-3,4-diol

(2R,3R,4R,5R)-1-butyl-2- (hydroxymethyl)-5-methylpyrrolidine- 3,4-diol

(2R,3R,4S,5R)-2-(hydroxymethyl)-5- methyl-1-nonylpyrrolidine-3,4-diol

(2R,3S,4S,5R)-2,5- dimethylpyrrolidine-3,4-diol

(2R,3R,4R,5R)-2,5- dimethylpyrrolidine-3,4-diol

(2R,3S,4R,5R)-2,5- dimethylpyrrolidine-3,4-diol

(2R,3R,4S,5R)-1-benzyl-2,5- dimethylpyrrolidine-3,4-diol

In alternative embodiments of the invention, compounds according toFormula (I) include one or more of the compounds described in Table 2 inwhich one or more OH groups are replaced by F or H.

In alternative embodiments of the invention, one or more of thecompounds described in Table 2 in which one or more OH groups isreplaced by F or H are specifically excluded from the compoundsdescribed in Formula (I).

As will be appreciated by a person skilled in the art, Formula (I) abovemay also be represented alternatively as follows:

As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. For example, “acompound” refers to one or more of such compounds, while “the enzyme”includes a particular enzyme as well as other family members andequivalents thereof as known to those skilled in the art.

Throughout this application, it is contemplated that the term “compound”or “compounds” refers to the compounds discussed herein and includesprecursors and derivatives of the compounds, including acyl-protectedderivatives, and pharmaceutically acceptable salts of the compounds,precursors, and derivatives. The invention also includes prodrugs of thecompounds, pharmaceutical compositions including the compounds and apharmaceutically acceptable carrier, and pharmaceutical compositionsincluding prodrugs of the compounds and a pharmaceutically acceptablecarrier.

The compounds of the present invention may contain one or moreasymmetric centers and can thus occur as racemates and racemic mixtures,single enantiomers, diastereomeric mixtures and individualdiastereomers. Additional asymmetric centers may be present dependingupon the nature of the various substituents on the molecule. Each suchasymmetric center will independently produce two optical isomers and itis intended that all of the possible optical isomers and diastereomersin mixtures and as pure or partially purified compounds are includedwithin the ambit of this invention. Any formulas, structures or names ofcompounds described in this specification that do not specify aparticular stereochemistry are meant to encompass any and all existingisomers as described above and mixtures thereof in any proportion. Whenstereochemistry is specified, the invention is meant to encompass thatparticular isomer in pure form or as part of a mixture with otherisomers in any proportion.

“Alkyl” refers to a straight or branched hydrocarbon chain groupconsisting solely of carbon and hydrogen atoms, containing nounsaturation and including, for example, from one to eighteen carbonatoms, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, or 18 carbon atoms, and which is attached to the rest of themolecule by a single bond. In alternative embodiments, the alkyl groupmay contain from one to ten carbon atoms, such as 1, 2, 3, 4, 5, 6, 7,8, 9, or 10 carbon atoms. In alternative embodiments, the alkyl groupmay contain from one to eight carbon atoms, such as 1, 2, 3, 4, 5, 6, 7,or 8 carbon atoms. In alternative embodiments, the alkyl group maycontain from one to six carbon atoms, such as 1, 2, 3, 4, 5, or 6 carbonatoms. Unless stated otherwise specifically in the specification, thealkyl group may be optionally substituted by one or more substituents asdescribed herein. Unless stated otherwise specifically herein, it isunderstood that the substitution can occur on any carbon of the alkylgroup.

“Alkenyl” refers to a straight or branched hydrocarbon chain groupconsisting solely of carbon and hydrogen atoms, containing at least onedouble bond and including, for example, from two to ten carbon atoms,such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, and which isattached to the rest of the molecule by a single bond or a double bond.In alternative embodiments, the alkenyl group may contain from two toeight carbon atoms, such as 2, 3, 4, 5, 6, 7, or 8 carbon atoms. Inalternative embodiments, the alkenyl group may contain from three to sixcarbon atoms, such as 3, 4, 5, or 6 carbon atoms. Unless statedotherwise specifically in the specification, the alkenyl group may beoptionally substituted by one or more substituents as described herein.Unless stated otherwise specifically herein, it is understood that thesubstitution can occur on any carbon of the alkenyl group.

“Alkynyl” refers to a straight or branched hydrocarbon chain groupconsisting solely of carbon and hydrogen atoms, containing at least onetriple bond and including, for example, from two to ten carbon atoms. Inalternative embodiments, the alkynyl group may contain from two to eightcarbon atoms, such as 2, 3, 4, 5, 6, 7, or 8 carbon atoms. Inalternative embodiments, the alkynyl group may contain from three to sixcarbon atoms, such as 3, 4, 5, or 6 carbon atoms. Unless statedotherwise specifically in the specification, the alkynyl group may beoptionally substituted by one or more substituents as described herein.

“Aryl” refers to a mono- or bicyclic aromatic ring containing onlycarbon atoms, including for example, 6-14 members, such as 6, 7, 8, 9,10, 11, 12, 13, or 14 members. Examples of aryl groups include phenyl,biphenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl,2,3-dihydrobenzofuranyl, dihydrobenzopyranyl, 1,4-benzodioxanyl, and thelike. Unless stated otherwise specifically herein, the term “aryl” ismeant to include aryl groups optionally substituted by one or moresubstituents as described herein.

“Heteroaryl” refers to a single or fused aromatic ring group containingone or more heteroatoms in the ring, for example N, O, S, including forexample, 5-14 members, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14members. Examples of heteroaryl groups include furan, thiophene,pyrrole, oxazole, thiazole, imidazole, pyrazole, isoxazole, isothiazole,1,2,3-oxadiazole, 1,2,3-triazole, 1,2,4-triazole, 1,3,4-thiadiazole,tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, 1,3,5-triazine,imidazole, benzimidazole, benzoxazole, benzothiazole, indolizine,indole, isoindole, benzofuran, benzothiophene, 1H-indazole, purine,4H-quinolizine, quinoline, isoquinoline, cinnoline, phthalazine,quinazoline, quinoxaline, 1,8-naphthyridine, pteridine, and the like.Unless stated otherwise specifically herein, the term “heteroaryl” ismeant to include heteroaryl groups optionally substituted by one or moresubstituents as described herein.

“Arylalkyl” refers to a group of the formula —R_(a)R_(b) where R_(a) isa C₁₋₁₀ alkyl group as described herein and R_(b) is one or more arylmoieties as described herein. The aryl group(s) may be optionallysubstituted as described herein.

“Arylalkenyl” refers to a group of the formula —R_(c)R_(b) where R_(c)is a C₂₋₁₀ alkenyl group as described herein and R_(b) is one or morearyl moieties as described herein. The aryl group(s) may be optionallysubstituted as described herein.

“Arylalkynyl” refers to a group of the formula —R_(d)R_(b) where R_(d)is a C₂₋₁₀ alkynyl group as described herein and R_(b) is one or morearyl moieties as described herein. The aryl group(s) may be optionallysubstituted as described herein.

“Heteroarylalkyl” refers to a group of the formula —R_(a)R_(c) whereR_(a) is a C₁₋₁₀ alkyl group as described herein and R_(c) is one ormore heteroaryl moieties as described herein. The aryl group(s) may beoptionally substituted as described herein.

“Heteroarylalkenyl” refers to a group of the formula —R_(c)R_(e) whereR_(c) is a C₂₋₁₀ alkenyl group as described herein and R_(e) is one ormore heteroaryl moieties as described herein. The heteroaryl group(s)may be optionally substituted as described herein.

“Heteroarylalkynyl” refers to a group of the formula —R_(d)R_(e) whereR_(d) is a C₂₋₁₀ alkynyl group as described herein and R_(e) is one ormore heteroaryl moieties as described herein. The heteroaryl group(s)may be optionally substituted as described herein.

“Acyl” refers to a group of the formula —C(O)R_(f), where R_(f) is H ora C₁₋₁₀ alkyl group or a C₁₋₆ alkyl group or a C₃₋₁₅ cycloalkyl group asdescribed herein. The alkyl or cycloalkyl group(s) may be optionallysubstituted as described herein.

“Arylalkylacyl” refers to a group of the formula —C(O)R_(g)R_(b), whereR_(g) is a C₁₋₁₀ alkyl or a C₁₋₆ alkyl group as described herein andR_(b) is one or more aryl moieties as described herein. The alkyl oraryl group(s) may be optionally substituted as described herein.

“Heteroarylalkylacyl” refers to a group of the formula —C(O)R_(g)R_(e),where R_(g) is a C₁₋₁₀ alkyl or a C₁₋₆ alkyl group as described hereinand R_(e) is one or more heteroaryl moieties as described herein. Thealkyl or heteroaryl group(s) may be optionally substituted as describedherein.

“Alkoxy” refers to a group of the formula —OR_(g), where R_(g) is aC₁₋₁₀ alkyl or a C₁₋₆ alkyl group as described herein. The alkylgroup(s) may be optionally substituted as described herein.

“Cycloalkyl” refers to a stable monovalent monocyclic, bicyclic ortricyclic hydrocarbon group consisting solely of carbon and hydrogenatoms, having for example from 3 to 15 carbon atoms, and which issaturated and attached to the rest of the molecule by a single bond. Inalternative embodiments, the cycloalkyl group may contain from three tosix carbon atoms, such as 3, 4, 5, or 6 carbon atoms. Unless otherwisestated specifically herein, the term “cycloalkyl” is meant to includecycloalkyl groups which are optionally substituted as described herein.

“Halo” refers to bromo, chloro, fluoro, iodo, etc. In some embodiments,suitable halogens include fluorine or chlorine.

“Optional” or “optionally” means that the subsequently described eventof circumstances may or may not occur, and that the description includesinstances where said event or circumstance occurs one or more times andinstances in which it does not. For example, “optionally substitutedalkyl” means that the alkyl group may or may not be substituted and thatthe description includes both substituted alkyl groups and alkyl groupshaving no substitution, and that said alkyl groups may be substitutedone or more times. Examples of optionally substituted alkyl groupsinclude, without limitation, methyl, ethyl, propyl, etc. and includingcycloalkyls such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, etc.; examples of optionally substituted alkenyl groupsinclude allyl, crotyl, 2-pentenyl, 3-hexenyl, 2-cyclopentenyl,2-cyclohexenyl, 2-cyclopentenylmethyl, 2-cyclohexenylmethyl, etc. Insome embodiments, optionally substituted alkyl and alkenyl groupsinclude C₁₋₆ alkyls or alkenyls.

Therapeutic Indications

The invention provides methods of treating conditions that aremodulated, directly or indirectly, by an O-GlcNAcase enzyme or byO-GlcNAc-modified protein levels, for example, a condition that isbenefited by inhibition of an O-GlcNAcase enzyme or by an elevation ofO-GlcNAc-modified protein levels. Such conditions may include, withoutlimitation, Glaucoma, Schizophrenia, tauopathies, such as Alzheimer'sdisease, neurodegenerative diseases, cardiovascular diseases, diseasesassociated with inflammation, diseases associated with immunosuppressionand cancers. One or more of the compounds of the invention may also beuseful in the treatment of diseases or disorders related to deficiencyor over-expression of O-GlcNAcase or accumulation or depletion ofO-GlcNAc, or any disease or disorder responsive to glycosidaseinhibition therapy. Such diseases and disorders may include, but are notlimited to, Glaucoma, Schizophrenia, neurodegenerative disorders, suchas Alzheimer's disease (AD), or cancer. Such diseases and disorders mayalso include diseases or disorders related to the accumulation ordeficiency in the enzyme OGT. Also included is a method of protecting ortreating target cells expressing proteins that are modified by O-GlcNAcresidues, the dysregulation of which modification may result in diseaseor pathology. The term “treating” as used herein includes treatment,prevention, and amelioration.

In alternative embodiments, the invention provides methods of enhancingor elevating levels of protein O-GlcNAc modification in animal subjects,such as, veterinary and human subjects. This elevation of O-GlcNAclevels may be useful for the prevention or treatment of Alzheimer'sdisease; prevention or treatment of other neurodegenerative diseases(e.g. Parkinson's disease, Huntington's disease); providingneuroprotective effects; preventing damage to cardiac tissue; andtreating diseases associated with inflammation or immunosuppression.

In alternative embodiments, the invention provides methods ofselectively inhibiting an O-GlcNAcase enzyme in animal subjects, such asveterinary and human subjects.

In alternative embodiments, the invention provides methods of inhibitingphosphorylation of tau polypeptides, or inhibiting formation of NFTs, inanimal subjects, such as, veterinary and human subjects. Accordingly, acompound of the invention may be used to study and treat AD and othertauopathies.

In general, the methods of the invention may be effected byadministering a compound according to the invention to a subject in needthereof, or by contacting a cell or a sample with a compound accordingto the invention, for example, a pharmaceutical composition comprising atherapeutically effective amount of the compound according to Formula(I). More particularly, they may be useful in the treatment of adisorder in which the regulation of O-GlcNAc protein modification isimplicated, or any condition as described herein. Disease states ofinterest may include Alzheimer's disease (AD) and relatedneurodegenerative tauopathies, in which abnormal hyperphosphorylation ofthe microtubule-associated protein tau is involved in diseasepathogenesis. In some embodiments, a compound may be used to blockhyperphosphorylation of tau by maintaining elevated levels of O-GlcNAcon tau, thereby providing therapeutic benefit.

The effectiveness of a compound in treating pathology associated withthe accumulation of toxic tau species (for example, Alzheimer's diseaseand other tauopathies) may be confirmed by testing the ability of acompound to block the formation of toxic tau species in establishedcellular¹²¹⁻¹²³ and/or transgenic animal models of disease.^(33,34)

Tauopathies that may be treated with a compound of the invention mayinclude, without limitation: Alzheimer's disease, Amyotrophic lateralsclerosis (ALS), Amyotrophic lateral sclerosis with cognitive impairment(ALSci), Argyrophilic grain dementia, Bluit disease, Corticobasaldegeneration (CBD), Dementia pugilistica, Diffuse neurofibrillarytangles with calcification, Down's syndrome, Familial British dementia,Familial Danish dementia, Frontotemporal dementia with parkinsonismlinked to chromosome 17 (FTDP-17), Gerstmann-Straussler-Scheinkerdisease, Guadeloupean parkinsonism, Hallevorden-Spatz disease(neurodegeneration with brain iron accumulation type 1), Multiple systematrophy, Myotonic dystrophy, Niemann-Pick disease (type C),Pallido-ponto-nigral degeneration, Parkinsonism-dementia complex ofGuam, Pick's disease (PiD), Post-encephalitic parkinsonism (PEP), Priondiseases (including Creutzfeldt-Jakob Disease (CJD), VariantCreutzfeldt-Jakob Disease (vCJD), Fatal Familial Insomnia, and Kuru),Progressive supercortical gliosis, Progressive supranuclear palsy (PSP),Richardson's syndrome, Subacute sclerosing panencephalitis, Tangle-onlydementia, and Glaucoma.

One or more of the compounds of this invention may also be useful in thetreatment of conditions associate with tissue damage or stress,stimulating cells, or promoting differentiation of cells. Accordingly,in some embodiments, a compound of this invention may be used to providetherapeutic benefit in a variety of conditions or medical proceduresinvolving stress in cardiac tissue; such conditions may include, withoutlimitation: ischemia; hemorrhage; hypovolemic shock; myocardialinfarction; an interventional cardiology procedure; cardiac bypasssurgery; fibrinolytic therapy; angioplasty; and stent placement.

The effectiveness of a compound in treating pathology associated withcellular stress (including ischemia, hemorrhage, hypovolemic shock,myocardial infarction, and other cardiovascular disorders) may beconfirmed by testing the ability of a compound to prevent cellulardamage in established cellular stress assays,^(108,119,120) and toprevent tissue damage and promote functional recovery in animal modelsof ischemia-reperfusion,^(71,117) and trauma-hemorrhage.^(73,115,118)

Compounds that selectively inhibit O-GlcNAcase activity may be used forthe treatment of diseases that are associated with inflammation; suchconditions may include, without limitation: inflammatory or allergicdiseases such as asthma, allergic rhinitis, hypersensitivity lungdiseases, hypersensitivity pneumonitis, eosinophilic pneumonias,delayed-type hypersensitivity, atherosclerosis, interstitial lungdisease (ILD) (e.g., idiopathic pulmonary fibrosis, or ILD associatedwith rheumatoid arthritis, systemic lupus erythematosus, ankylosingspondylitis, systemic sclerosis, Sjogren's syndrome, polymyositis ordermatomyositis); systemic anaphylaxis or hypersensitivity responses,drug allergies, insect sting allergies; autoimmune diseases, such asrheumatoid arthritis, psoriatic arthritis, multiple sclerosis,Guillain-Barré syndrome, systemic lupus erythematosus, myastenia gravis,glomerulonephritis, autoimmune thyroiditis, graft rejection, includingallograft rejection or graft-versus-host disease; inflammatory boweldiseases, such as Crohn's disease and ulcerative colitis;spondyloarthropathies; scleroderma; psoriasis (including T-cell mediatedpsoriasis) and inflammatory dermatoses such as dermatitis, eczema,atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis(e.g., necrotizing, cutaneous, and hypersensitivity vasculitis);eosinphilic myotis, eosiniphilic fasciitis; and cancers.

In addition, compounds that affect levels of protein O-GlcNAcmodification may be used for the treatment of diseases associated withimmunosuppression, such as, for example, in individuals undergoingchemotherapy, radiation therapy, enhanced wound healing and burntreatment, therapy for autoimmune disease or other drug therapy (e.g.,corticosteroid therapy) or combination of conventional drugs used in thetreatment of autoimmune diseases and graft/transplantation rejection,which causes immunosuppression; or immunosuppression due to congenitaldeficiency in receptor function or other causes.

One or more of the compounds of the invention may be useful fortreatment of neurodegenerative diseases; such conditions may include,without limitation, Parkinson's disease and Huntington's disease. Otherconditions that may be treated are those triggered, affected, or in anyother way correlated with levels of O-GlcNAc post-translational proteinmodification. It is expected that one or more of the compounds of thisinvention may be useful for the treatment of such conditions and inparticular, but not limited to, the following for which a associationwith O-GlcNAc levels on proteins has been established: graft rejection,in particular but not limited to solid organ transplants, such as heart,lung, liver, kidney, and pancreas transplants (e.g. kidney and lungallografts); cancer, in particular but not limited to cancer of thebreast, lung, prostate, pancreas, colon, rectum, bladder, kidney, ovary;as well as non-Hodgkin's lymphoma and melanoma; epilepsy, pain,fibromyalgia, or stroke, e.g., for neuroprotection following a stroke.

Pharmaceutical & Veterinary Compositions, Dosages, and Administration

Pharmaceutical compositions including compounds according to theinvention, or for use according to the invention, are contemplated asbeing within the scope of the invention. In some embodiments,pharmaceutical compositions including an effective amount of a compoundof Formula (I) are provided.

The compounds of Formula (I) and their pharmaceutically acceptablesalts, enantiomers, solvates, and derivatives may be useful because theymay have pharmacological activity in animals, including humans. In someembodiments, one or more of the compounds according to the invention maybe stable in plasma, when administered to a subject.

In some embodiments, a compound according to the invention, or for useaccording to the invention, may be provided in combination with anyother active agents or pharmaceutical compositions where such combinedtherapy may be useful to modulate O-GlcNAcase activity, for example, totreat neurodegenerative, inflammatory, cardiovascular, orimmunoregulatory diseases, or any condition described herein. In someembodiments, a compound according to the invention, or for use accordingto the invention, may be provided in combination with one or more agentsuseful in the prevention or treatment of Alzheimer's disease. Examplesof such agents may include, without limitation,

-   -   acetylcholine esterase inhibitors (AChEIs) such as Aricept®        (Donepezil), Exelon® (Rivastigmine), Razadyne® (Razadyne ER®,        Reminyl®, Nivalin®, Galantamine), Cognex® (Tacrine), Dimebon,        Huperzine A, Phenserine, Debio-9902 SR (ZT-1 SR), Zanapezil        (TAK0147), ganstigmine, NP7557, etc.;    -   NMDA receptor antagonists such as Namenda® (Axura®, Akatinol®,        Ebixa®, Memantine), Dimebon, SGS-742, Neramexane, Debio-9902 SR        (ZT-1 SR), etc.;    -   gamma-secretase inhibitors and/or modulators such as Flurizan™        (Tarenflurbil, MPC-7869, R-flurbiprofen), LY450139, MK 0752,        E2101, BMS-289948, BMS-299897, BMS-433796, LY-411575, GSI-136,        etc.;    -   beta-secretase inhibitors such as ATG-Z1, CTS-21166, MK-8931,        etc.;    -   alpha-secretase activators, such as NGX267, etc;    -   amyloid-β aggregation and/or fibrillization inhibitors such as        Alzhemed™ (3APS, Tramiprosate, 3-amino-1-propanesulfonic acid),        AL-108, AL-208, AZD-103, PBT2, Cereact, ONO-2506PO, PPI-558,        etc.;    -   tau aggregation inhibitors such as methylene blue, etc.;    -   microtubule stabilizers such as AL-108, AL-208, paclitaxel,        etc.;    -   RAGE inhibitors, such as TTP488, etc.;    -   5-HT1a receptor antagonists, such as Xaliproden, Lecozotan,        etc.;    -   5-HT4 receptor antagonists, such as PRX-03410, etc.;    -   kinase inhibitors such as SRN-003-556, amfurindamide, LiCl,        AZD1080, NP031112, SAR-502250, etc.    -   humanized monoclonal anti-Aβ antibodies such as Bapineuzumab        (AAB-001), LY2062430, RN1219, ACU-5A5, etc.;    -   amyloid vaccines such as AN-1792, ACC-001, etc.;    -   neuroprotective agents such as Cerebrolysin, AL-108, AL-208,        Huperzine A, etc.;    -   L-type calcium channel antagonists such as MEM-1003, etc.;    -   nicotinic receptor antagonists, such as AZD3480, GTS-21, etc.;    -   nicotinic receptor agonists, such as MEM 3454, Nefiracetam,        etc.;    -   peroxisome proliferator-activated receptor (PPAR) gamma agonists        such as Avandia® (Rosglitazone), etc.;    -   phosphodiesterase IV (PDE4) inhibitors, such as MK-0952, etc.;    -   hormone replacement therapy such as estrogen (Premarin), etc.;    -   monoamine oxidase (MAO) inhibitors such as NS2330, Rasagiline        (Azilect®), TVP-1012, etc.;    -   AMPA receptor modulators such as Ampalex (CX 516), etc.;    -   nerve growth factors or NGF potentiators, such as CERE-110        (AAV-NGF), T-588, T-817MA, etc.;    -   agents that prevent the release of luteinizing hormone (LH) by        the pituitary gland, such as leuoprolide (VP-4896), etc.;    -   GABA receptor modulators such as AC-3933, NGD 97-1, CP-457920,        etc.;    -   benzodiazepine receptor inverse agonists such as SB-737552        (S-8510), AC-3933, etc.;    -   noradrenaline-releasing agents such as T-588, T-817MA, etc.

It is to be understood that combination of compounds according to theinvention, or for use according to the invention, with Alzheimer'sagents is not limited to the examples described herein, but may includecombination with any agent useful for the treatment of Alzheimer'sdisease. Combination of compounds according to the invention, or for useaccording to the invention, and other Alzheimer's agents may beadministered separately or in conjunction. The administration of oneagent may be prior to, concurrent to, or subsequent to theadministration of other agent(s).

In alternative embodiments, a compound may be supplied as a “prodrug” orprotected forms, which release the compound after administration to asubject. For example, a compound may carry a protective group which issplit off by hydrolysis in body fluids, e.g., in the bloodstream, thusreleasing the active compound or is oxidized or reduced in body fluidsto release the compound. Accordingly, a “prodrug” is meant to indicate acompound that may be converted under physiological conditions or bysolvolysis to a biologically active compound of the invention. Thus, theterm “prodrug” refers to a metabolic precursor of a compound of theinvention that is pharmaceutically acceptable. A prodrug may be inactivewhen administered to a subject in need thereof, but may be converted invivo to an active compound of the invention. Prodrugs are typicallyrapidly transformed in vivo to yield the parent compound of theinvention, for example, by hydrolysis in blood. The prodrug compoundoften offers advantages of solubility, tissue compatibility or delayedrelease in a subject.

The term “prodrug” is also meant to include any covalently bondedcarriers which release the active compound of the invention in vivo whensuch prodrug is administered to a subj ect. Prodrugs of a compound ofthe invention may be prepared by modifying functional groups present inthe compound of the invention in such a way that the modifications arecleaved, either in routine manipulation or in vivo, to the parentcompound of the invention. Prodrugs include compounds of the inventionwhere a hydroxy, amino or mercapto group is bonded to any group that,when the prodrug of the compound of the invention is administered to amammalian subject, cleaves to form a free hydroxy, free amino or freemercapto group, respectively. Examples of prodrugs include, but are notlimited to, acetate, formate and benzoate derivatives of alcohol andacetamide, formamide, and benzamide derivatives of amine functionalgroups in one or more of the compounds of the invention and the like.

A discussion of prodrugs may be found in “Smith and Williams'Introduction to the Principles of Drug Design,” H. J. Smith, Wright,Second Edition, London (1988); Bundgard, H., Design of Prodrugs (1985),pp. 7-9, 21-24 (Elsevier, Amsterdam); The Practice of MedicinalChemistry, Camille G. Wermuth et al., Ch 31, (Academic Press, 1996); ATextbook of Drug Design and Development, P. Krogsgaard-Larson and H.Bundgaard, eds. Ch 5, pgs 113 191 (Harwood Academic Publishers, 1991);Higuchi, T., et al., “Pro-drugs as Novel Delivery Systems,” A.C.S.Symposium Series, Vol. 14; or in Bioreversible Carriers in Drug Design,ed. Edward B. Roche, American Pharmaceutical Association and PergamonPress, 1987.

Suitable prodrug forms of one or more of the compounds of the inventionmay include embodiments in which one or more OH groups as set forth inFormula (I) may be protected as OC(O)R, where R may be optionallysubstituted alkyl, alkenyl, alkynyl, aryl, or heteroaryl. In these casesthe ester groups may be hydrolyzed in vivo (e.g. in bodily fluids),liberating the OH groups and releasing the active compounds. Preferredprodrug embodiments of the invention may include compounds of Formula(I) where one or more OH groups may be protected with acetate, forexample as OC(O)CH₃.

Compounds according to the invention, or for use according to theinvention, may be provided alone or in combination with other compoundsin the presence of a liposome, an adjuvant, or any pharmaceuticallyacceptable carrier, diluent or excipient, in a form suitable foradministration to a subject such as a mammal, for example, humans,cattle, sheep, etc. If desired, treatment with a compound according tothe invention may be combined with more traditional and existingtherapies for the therapeutic indications described herein. Compoundsaccording to the invention may be provided chronically orintermittently. “Chronic” administration refers to administration of thecompound(s) in a continuous mode as opposed to an acute mode, so as tomaintain the initial therapeutic effect (activity) for an extendedperiod of time. “Intermittent” administration is treatment that is notconsecutively done without interruption, but rather is cyclic in nature.The terms “administration,” “administrable,” or “administering” as usedherein should be understood to mean providing a compound of theinvention to the subject in need of treatment.

“Pharmaceutically acceptable carrier, diluent or excipient” may include,without limitation, any adjuvant, carrier, excipient, glidant,sweetening agent, diluent, preservative, dye/colorant, flavor enhancer,surfactant, wetting agent, dispersing agent, suspending agent,stabilizer, isotonic agent, solvent, or emulsifier that has beenapproved, for example, by the United States Food and Drug Administrationor other governmental agency as being acceptable for use in humans ordomestic animals.

A compound of the present invention may be administered in the form of apharmaceutically acceptable salt. In such cases, pharmaceuticalcompositions in accordance with this invention may comprise a salt ofsuch a compound, preferably a physiologically acceptable salt, which areknown in the art. In some embodiments, the term “pharmaceuticallyacceptable salt” as used herein means an active ingredient comprisingcompounds of Formula I used in the form of a salt thereof, particularlywhere the salt form confers on the active ingredient improvedpharmacokinetic properties as compared to the free form of the activeingredient or other previously disclosed salt form.

A “pharmaceutically acceptable salt” may include both acid and baseaddition salts. A “pharmaceutically acceptable acid addition salt”refers to those salts which retain the biological effectiveness andproperties of the free bases, which are not biologically or otherwiseundesirable, and which may be formed with inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like, and organic acids such as acetic acid,trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like.

A “pharmaceutically acceptable base addition salt” refers to those saltswhich may retain the biological effectiveness and properties of the freeacids, which may not be biologically or otherwise undesirable. Thesesalts may be prepared from addition of an inorganic base or an organicbase to the free acid. Salts derived from inorganic bases may include,but are not limited to, the sodium, potassium, lithium, ammonium,calcium, magnesium, iron, zinc, copper, manganese, aluminum salts andthe like. Preferred inorganic salts may be the ammonium, sodium,potassium, calcium, and magnesium salts. Salts derived from organicbases may include, but are not limited to, salts of primary, secondary,and tertiary amines, substituted amines including naturally occurringsubstituted amines, cyclic amines and basic ion exchange resins, such asisopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, 2-dimethylaminoethanol,2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine,caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,glucosamine, methylglucamine, theobromine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like.Particularly preferred organic bases may be isopropylamine,diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, cholineand caffeine.

Thus, the term “pharmaceutically acceptable salt” encompasses allacceptable salts including but not limited to acetate, lactobionate,benzenesulfonate, laurate, benzoate, malate, bicarbonate, maleate,bisulfate, mandelate, bitartarate, mesylate, borate, methylbromide,bromide, methylnitrite, calcium edetate, methylsulfate, camsylate,mucate, carbonate, napsylate, chloride, nitrate, clavulanate,N-methylglucamine, citrate, ammonium salt, dihydrochloride, oleate,edetate, oxalate, edisylate, pamoate (embonate), estolate, palmitate,esylate, pantothenate, fumarate, phosphate/diphosphate, gluceptate,polygalacturonate, gluconate, salicylate, glutame, stearate,glycollylarsanilate, sulfate, hexylresorcinate, subacetate, hydradamine,succinate, hydrobromide, tannate, hydrochloride, tartrate,hydroxynaphthoate, teoclate, iodide, tosylate, isothionate,triethiodide, lactate, panoate, valerate, and the like.

Pharmaceutically acceptable salts of a compound of the present inventionmay be used as a dosage for modifying solubility or hydrolysischaracteristics, or may be used in sustained release or prodrugformulations. Also, pharmaceutically acceptable salts of a compound ofthis invention may include those formed from cations such as sodium,potassium, aluminum, calcium, lithium, magnesium, zinc, and from basessuch as ammonia, ethylenediamine, N-methyl-glutamine, lysine, arginine,omithine, choline, N,N′-dibenzylethylene-diamine, chloroprocaine,diethanolamine, procaine, N-benzylphenethyl-amine, diethylamine,piperazine, tris(hydroxymethyl)aminomethane, and tetramethylammoniumhydroxide.

Pharmaceutical formulations may typically include one or more carriersacceptable for the mode of administration of the preparation, be it byinjection, inhalation, topical administration, lavage, or other modessuitable for the selected treatment. Suitable carriers may be thoseknown in the art for use in such modes of administration.

Suitable pharmaceutical compositions may be formulated by means known inthe art and their mode of administration and dose determined by theskilled practitioner. For parenteral administration, a compound may bedissolved in sterile water or saline or a pharmaceutically acceptablevehicle used for administration of non-water soluble compounds such asthose used for vitamin K. For enteral administration, the compound maybe administered in a tablet, capsule or dissolved in liquid form. Thetable or capsule may be enteric coated, or in a formulation forsustained release. Many suitable formulations are known, including,polymeric or protein microparticles encapsulating a compound to bereleased, ointments, gels, hydrogels, or solutions which can be usedtopically or locally to administer a compound. A sustained release patchor implant may be employed to provide release over a prolonged period oftime. Many techniques known to skilled practitioners are described inRemington: the Science & Practice of Pharmacy by Alfonso Gennaro,20^(th) ed., Williams & Wilkins, (2000). Formulations for parenteraladministration may, for example, contain excipients, polyalkyleneglycols such as polyethylene glycol, oils of vegetable origin, orhydrogenated naphthalenes. Biocompatible, biodegradable lactide polymer,lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylenecopolymers may be used to control the release of a compound. Otherpotentially useful parenteral delivery systems for modulatory compoundsmay include ethylene-vinyl acetate copolymer particles, osmotic pumps,implantable infusion systems, and liposomes. Formulations for inhalationmay contain excipients, for example, lactose, or may be aqueoussolutions containing, for example, polyoxyethylene-9-lauryl ether,glycocholate and deoxycholate, or may be oily solutions foradministration in the form of nasal drops, or as a gel.

A compound or a pharmaceutical composition according to the presentinvention may be administered by oral or non-oral, e.g., intramuscular,intraperitoneal, intravenous, intracisternal injection or infusion,subcutaneous injection, transdermal or transmucosal routes. In someembodiments, a compound or pharmaceutical composition in accordance withthis invention or for use in this invention may be administered by meansof a medical device or appliance such as an implant, graft, prosthesis,stent, etc. Implants may be devised which are intended to contain andrelease such compounds or compositions. An example would be an implantmade of a polymeric material adapted to release the compound over aperiod of time. A compound may be administered alone or as a mixturewith a pharmaceutically acceptable carrier e.g., as solid formulationssuch as tablets, capsules, granules, powders, etc.; liquid formulationssuch as syrups, injections, etc.; injections, drops, suppositories,pessaryies. In some embodiments, compounds or pharmaceuticalcompositions in accordance with this invention or for use in thisinvention may be administered by inhalation spray, nasal, vaginal,rectal, sublingual, or topical routes and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration.

A compound of the invention may be used to treat animals, includingmice, rats, horses, cattle, sheep, dogs, cats, and monkeys. However, acompound of the invention may also be used in other organisms, such asavian species (e.g., chickens). One or more of the compounds of theinvention may also be effective for use in humans. The term “subject” oralternatively referred to herein as “patient” is intended to be referredto an animal, preferably a mammal, most preferably a human, who has beenthe object of treatment, observation or experiment. However, one or moreof the compounds, methods and pharmaceutical compositions of the presentinvention may be used in the treatment of animals. Accordingly, as usedherein, a “subject” may be a human, non-human primate, rat, mouse, cow,horse, pig, sheep, goat, dog, cat, etc. The subject may be suspected ofhaving or at risk for having a condition that may require modulation ofO-GlcNAcase activity.

An “effective amount” of a compound according to the invention mayinclude a therapeutically effective amount or a prophylacticallyeffective amount. A “therapeutically effective amount” refers to anamount effective, at dosages and for periods of time necessary, toachieve the desired therapeutic result, such as inhibition of anO-GlcNAcase, elevation of O-GlcNAc levels, inhibition of tauphosphorylation, or any condition described herein. A therapeuticallyeffective amount of a compound may vary according to factors such as thedisease state, age, sex, and weight of the individual, and the abilityof the compound to elicit a desired response in the individual. Dosageregimens may be adjusted to provide the optimum therapeutic response. Atherapeutically effective amount may also be one in which any toxic ordetrimental effects of the compound are outweighed by thetherapeutically beneficial effects. A “prophylactically effectiveamount” may refer to an amount effective, at dosages and for periods oftime necessary, to achieve the desired prophylactic result, such asinhibition of an O-GlcNAcase, elevation of O-GlcNAc levels, inhibitionof tau phosphorylation, or any condition described herein. Typically, aprophylactic dose may be used in subjects prior to or at an earlierstage of disease, so that a prophylactically effective amount may beless than a therapeutically effective amount. A suitable range fortherapeutically or prophylactically effective amounts of a compound maybe any integer from 0.1 nM-0.1 M, 0.1 nM-0.05 M, 0.05 nM-15 M or 0.01nM-10 μM.

In alternative embodiments, in the treatment or prevention of conditionswhich may require modulation of O-GlcNAcase activity, an appropriatedosage level may generally be about 0.01 to 500 mg per kg subject bodyweight per day, and may be administered in singe or multiple doses. Insome embodiments, the dosage level may be about 0.1 to about 250 mg/kgper day. It will be understood that the specific dose level andfrequency of dosage for any particular patient may be varied and maydepend upon a variety of factors including the activity of the specificcompound used, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the patient undergoing therapy.

It is to be noted that dosage values may vary with the severity of thecondition to be alleviated. For any particular subject, specific dosageregimens may be adjusted over time according to the individual need andthe professional judgement of the person administering or supervisingthe administration of the compositions. Dosage ranges set forth hereinare exemplary only and do not limit the dosage ranges that may beselected by medical practitioners. The amount of active compound(s) inthe composition may vary according to factors such as the disease state,age, sex, and weight of the subject. Dosage regimens may be adjusted toprovide the optimum therapeutic response. For example, a single bolusmay be administered, several divided doses may be administered over timeor the dose may be proportionally reduced or increased as indicated bythe exigencies of the therapeutic situation. It may be advantageous toformulate parenteral compositions in dosage unit form for ease ofadministration and uniformity of dosage. In general, compounds of theinvention should be used without causing substantial toxicity, and asdescribed herein, one or more of the compounds may exhibit a suitablesafety profile for therapeutic use. Toxicity of a compound of theinvention may be determined using standard techniques, for example, bytesting in cell cultures or experimental animals and determining thetherapeutic index, i.e., the ratio between the LD50 (the dose lethal to50% of the population) and the LD100 (the dose lethal to 100% of thepopulation). In some circumstances however, such as in severe diseaseconditions, it may be necessary to administer substantial excesses ofthe compositions.

In the compounds of generic Formula (I), the atoms may exhibit theirnatural isotopic abundances, or one or more of the atoms may beartificially enriched in a particular isotope having the same atomicnumber, but an atomic mass or mass number different from the atomic massor mass number predominantly found in nature. The present invention ismeant to include all suitable isotopic variations of the compounds ofgeneric Formula (I). For example, different isotopic forms of hydrogen(H) include protium (¹H), deuterium (²H) and tritium (³H). Protium isthe predominant hydrogen isotope found in nature. Enriching fordeuterium may afford certain therapeutic advantages, such as increasingin vivo half-life or reducing dosage requirements, or may provide acompound useful as a standard for characterization of biologicalsamples. Isotopically-enriched compounds within generic Formula (I) maybe prepared without undue experimentation by conventional techniqueswell known to those skilled in the art or by processes analogous tothose described in the Schemes and Examples herein using appropriateisotopically-enriched reagents and/or intermediates.

Other Uses and Assays

A compound of Formula (I) may be used in screening assays for compoundswhich modulate the activity of glycosidase enzymes, preferably theO-GlcNAcase enzyme. The ability of a test compound to inhibitO-GlcNAcase-dependent cleavage of O-GlcNAc from a model substrate may bemeasured using any assays, as described herein or known to one ofordinary skill in the art. For example, a fluoresence or UV-based assayknown in the art may be used. A “test compound” may be anynaturally-occurring or artificially-derived chemical compound. Testcompounds may include, without limitation, peptides, polypeptides,synthesised organic molecules, naturally occurring organic molecules,and nucleic acid molecules. A test compound may “compete” with a knowncompound such as a compound of Formula (I) by, for example, interferingwith inhibition of O-GlcNAcase-dependent cleavage of O-GlcNAc or byinterfering with any biological response induced by a compound ofFormula (I).

Generally, a test compound may exhibit any value between 10% and 200%,or over 500%, modulation when compared to a compound of Formula (I) orother reference compound. For example, a test compound may exhibit atleast any positive or negative integer from 10% to 200% modulation, orat least any positive or negative integer from 30% to 150% modulation,or at least any positive or negative integer from 60% to 100%modulation, or any positive or negative integer over 100% modulation. Acompound that is a negative modulator may in general decrease modulationrelative to a known compound, while a compound that is a positivemodulator may in general increase modulation relative to a knowncompound.

In general, test compounds may be identified from large libraries ofboth natural products or synthetic (or semi-synthetic) extracts orchemical libraries according to methods known in the art. Those skilledin the field of drug discovery and development will understand that theprecise source of test extracts or compounds is not critical to themethod(s) of the invention. Accordingly, virtually any number ofchemical extracts or compounds may be screened using the exemplarymethods described herein. Examples of such extracts or compounds mayinclude, but are not limited to, plant-, fungal-, prokaryotic- oranimal-based extracts, fermentation broths, and synthetic compounds, aswell as modification of existing compounds. Numerous methods are alsoavailable for generating random or directed synthesis (e.g.,semi-synthesis or total synthesis) of any number of chemical compounds,that may include, without limitation, saccharide-, lipid-, peptide-, andnucleic acid-based compounds. Synthetic compound libraries arecommercially available. Alternatively, libraries of natural compounds inthe form of bacterial, fungal, plant, and animal extracts arecommercially available from a number of sources, including Biotics(Sussex, UK), Xenova (Slough, UK), Harbor Branch Oceanographic Institute(Ft. Pierce, Fla., USA), and PharmaMar, MA, USA. In addition, naturaland synthetically produced libraries may be produced, if desired,according to methods known in the art, e.g., by standard extraction andfractionation methods. Furthermore, if desired, any library or compoundmay be readily modified using standard chemical, physical, orbiochemical methods.

When a crude extract is found to modulate inhibition ofO-GlcNAcase-dependent cleavage of O-GlcNAc, or any biological responseinduced by a compound of Formula (I), further fractionation of thepositive lead extract may be necessary to isolate chemical constituentsresponsible for the observed effect. Thus, the goal of the extraction,fractionation, and purification process is the careful characterizationand identification of a chemical entity within the crude extract havingO-GlcNAcase-inhibitory activities. The same assays described herein forthe detection of activities in mixtures of compounds may be used topurify the active component and to test derivatives thereof. Methods offractionation and purification of such heterogeneous extracts are knownin the art. If desired, compounds shown to be useful agents fortreatment may be chemically modified according to methods known in theart. Compounds identified as being of therapeutic, prophylactic,diagnostic, or other value may be subsequently analyzed using a suitableanimal model, as described herein on known in the art.

In some embodiments, one or more of the compounds may be useful in thedevelopment of animal models for studying diseases or disorders that maybe related to deficiencies in O-GlcNAcase, over-expression ofO-GlcNAcase, accumulation of O-GlcNAc, depletion of O-GlcNAc, and forstudying treatment of diseases and disorders that may be related todeficiency or over-expression of O-GlcNAcase, or accumulation ordepletion of O-GlcNAc. Such diseases and disorders may includeneurodegenerative diseases, including Alzheimer's disease, and cancer.

Various alternative embodiments and examples of the invention aredescribed herein. These embodiments and examples are illustrative andshould not be construed as limiting the scope of the invention.

EXAMPLES

The following examples are intended to illustrate embodiments of theinvention and are not intended to be construed in a limiting manner.

Abbreviations

-   ABCN=1,1′-azobis(cyclohexane-carbonitrile)-   AcCl=acetyl chloride-   AIBN=azobisisobutyronitrile-   BCl₃=boron trichloride-   BnBr=benzyl bromide-   Bu₄NI=tetra-n-butylammonium iodide-   Boc₂O=di-tert-butyl dicarbonate-   BzC1=benzoyl chloride-   DAST=diethylaminosulfur trifluoride-   DCM=dichloromethane-   DIAD=diisopropyl azodicarboxylate-   DIPEA=diisopropylethylamine-   DMAP=4-dimethylaminopyridine-   DMF=N,N-dimethylformamide-   DMP=Dess-Martin periodinane-   DMSO=dimethyl sulfoxide-   EDCI=1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-   Et₃N=triethylamine-   Et₂O=diethyl ether-   HATU=(O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate)-   PMB=pentamethylbenzene-   TBDMSCl=tert-butyldimethylsilyl chloride-   TBAF=tetra-n-butylammonium fluoride-   TMSCF₃=(trifluoromethyl)trimethylsilane-   TFA=2,2,2-trifluoroacetic acid-   THF=tetrahydrofuran-   thio-CDI=1,1′-thiocarbonyl diimidazole    General Procedures and Intermediates

The compounds of the invention are synthesized according to standardschemes and procedures, as indicated in, for example, Schemes 1 to 27,as appropriate. Intermediate A (Scheme 15) may be prepared as describedin, for example, Tetrahedron 2009, 65, 93-100. Intermediate B (Scheme26) may be prepared as described in, for example, Tetrahedron 1994, 50,4215 and PCT WO 1997/009040. For Schemes 2 to 27, the general proceduresare defined as follows:

General procedure A: Pd(OH)₂/H₂, MeOH, 50° C., 50 psi

General Procedure B: RCHO, NaBH₃CN, HOAc, MeOH

General Procedure C: 4N HCl, 40° C.

General Procedure D: RCOOH, HATU, DMF, i-Pr₂NEt

General Procedure E: RBr, K₂CO₃, DMF

It is to be understood that any suitable scheme within the knowledge ofone skilled in the art may be used to synthesize one or more of thecompounds described herein.

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using literature procedures as indicated in Scheme 1(Tetrahedron Letters, 1997, 38(31), 5463-5466; J. CarbohydrateChemistry, 2000, 19(8), 975-990). Characterization data for its CHCOOHsalt: ¹H NMR (400 MHz, CD₃OD) δ 3.87-3.83 (m, 1H), 3.79-3.72 (m, 2H),3.67-3.63 (m, 1H), 3.49-3.44 (m, 1H), 3.42-3.38 (m, 1H), 3.26-3.18 (m,2H), 1.98 (s, 3H), 1.93 (CH₃COOH); ¹³C NMR (125 MHz, CD₃OD) δ 179.97,174.84, 78.61, 77.13, 65.37, 63.68, 60.47, 40.91, 23.82, 22.71. MS, (ES,m/z) [M+H]⁺ 205.13.

General Procedure for Reductive Amination

To a mixture ofN-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide(0.030 g, 0.15 mmol) and aldehyde (0.45 mmol) in methanol (2 mL) wasadded NaBH₃CN (0.030 g, 0.48 mmol). After the mixture was stirred atroom temperature overnight, the solvent was removed under reducedpressure, and the residue was purified on silica gel by flash columnchromatography (EtOAc, then MeOH/DCM, 1:8) to afford the product.

N-(((2R,3R,4R,5R)-1-benzyl-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-benzyl-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, usingbenzaldehyde. Very pale yellow oil. Yield 43%. ¹H NMR (400 MHz, CD₃OD) δ7.42-7.40 (m, 2H), 7.30-7.28 (m, 2H), 7.23-7.19 (m, 1H), 4.01 (t, J=2.4Hz, 1H), 3.98-3.93 (m, 2H), 3.83-3.82 (m, 1H), 3.77 (dd, J=4.9, 11.4 Hz,1H), 3.63 (dd, J=3.3, 11.4 Hz, 1H), 3.42 (dd, J=3.5, 13.9 Hz, 1H), 3.28(dd, J=6.0, 13.9 Hz, 1H), 3.14-3.11 (m, 1H), 3.05-3.02 (m, 1H), 1.92 (s,3H); ¹³C NMR (100 MHz, CD₃OD) δ 173.66, 140.90, 129.50, 129.46, 128.12,81.29, 80.80, 70.07, 69.93, 61.15, 52.49, 40.38, 22.81. MS, (ES, m/z)[M+H]⁺ 295.18.

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-phenethylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-phenethylpyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, usingphenylacetaldehyde. Very pale yellow oil. Yield 45%. ¹H NMR (400 MHz,CD₃OD) δ 7.29-7.24 (m, 4H), 7.19-7.14 (m, 1H), 3.97 (t, J=2.3 Hz, 1H),3.83 (dd, J=4.7, 11.4 Hz, 1H), 3.74-3.70 (m, 2H), 3.54-3.50 (m, 1H),3.17-3.14 (m, 1H), 3.11-3.02 (m, 3H), 2.89-3.83 (m, 2H), 2.79-2.74 (m,1H), 1.84 (s, 3H); ¹³C NMR (100 MHz, CD₃OD) δ 173.53, 142.19, 129.98,129.49, 127.20, 81.03, 80.96, 69.87, 69.36, 60.95, 49.70, 39.41, 35.88,22.89. MS, (ES, m/z) [M+H]⁺ 309.19.

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-propylpyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-propylpyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, usingpropanal. Colorless oil. Yield 53%. ¹H NMR (500 MHz, CD₃OD) δ 3.97-3.96(m, 1H), 3.78-3.74 (m, 1H), 3.76 (dd, J=5.1, 11.4 Hz, 1H), 3.68 (dd,J=3.4, 11.4 Hz, 1H), 3.50 (dd, J=3.3, 13.7 Hz, 1H), 3.19 (dd, J=7.0,13.7 Hz, 1H), 3.05-3.01 (m, 2H), 2.77-2.72 (m, 1H), 2.64-2.58 (m, 1H),1.95 (s, 3H), 1.65-1.57 (m, 1H), 1.56-1.48 (m, 1H), 0.94 (t, J=7.4 Hz,3H); ¹³C NMR (125 MHz, CD₃OD) δ 173.70, 81.21, 80.99, 70.36, 69.37,60.69, 50.08, 39.43, 22.78, 22.50, 12.35. MS, (ES, m/z) [M+H]⁺ 247.17.

N-(((2R,3R,4R,5R)-1-butyl-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-butyl-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, usingbutanal. Colorless oil. Yield 70%. ¹H NMR (500 MHz, CD₃OD) δ 3.99-3.98(m, 1H), 3.82-3.81 (m, 1H), 3.78 (dd, J=4.9, 11.5 Hz, 1H), 3.71 (dd,J=3.7, 11.5 Hz, 1H), 3.51 (dd, J=3.5, 13.8 Hz, 1H), 3.26 (dd, J=6.8,13.8 Hz, 1H), 3.14-3.08 (m, 2H), 2.84-2.77 (m, 2H), 1.96 (s, 3H),1.59-1.54 (m, 2H), 1.43-1.36 (m, 2H), 0.95 (t, J=7.4 Hz, 3H); ¹³C NMR(125 MHz, CD₃OD) δ 173.90, 80.90, 80.68, 70.79, 69.80, 60.57, 48.28,39.38, 31.21, 22.76, 21.66, 14.43. MS, (ES, m/z) [M+H]⁺ 261.17.

N-(((2R,3R,4R,5R)-1-hexyl-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-hexyl-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, usinghexanal. Colorless oil. Yield 73%. ¹H NMR (500 MHz, CD₃OD) δ 3.98-3.97(m, 1H), 3.82-3.81 (m, 1H), 3.78 (dd, J=4.9, 11.6 Hz, 1H), 3.71 (dd,J=3.8, 11.6 Hz, 1H), 3.51 (dd, J=3.6, 13.8 Hz, 1H), 3.26 (dd, J=6.8,13.8 Hz, 1H), 3.14-3.09 (m, 2H), 2.84-2.75 (m, 2H), 1.95 (s, 3H),1.62-1.55 (m, 2H), 1.38-1.28 (m, 6H), 0.91 (t, J=7.3 Hz, 3H); ¹³C NMR(125 MHz, CD₃OD) δ 173.93, 80.83, 80.62, 70.85, 69.88, 60.53, 48.63,39.38, 32.99, 28.97, 28.26, 23.87, 22.76, 14.54. MS, (ES, m/z) [M+H]⁺289.21.

EXAMPLES Example 1N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, usinghydrocinnamaldehyde. Very pale yellow oil. Yield 50%. ¹H NMR (400 MHz,CD₃OD) δ 7.26-7.20 (m, 4H), 7.16-7.13 (m, 1H), 3.98-3.97 (m, 1H),3.79-3.78 (m, 1H), 3.73 (dd, J=5.0, 11.3 Hz, 1H), 3.65 (dd, J=3.3, 11.3Hz, 1H), 3.48 (dd, J=3.5, 13.6 Hz, 1H), 3.17 (dd, J=7.0, 13.6 Hz, 1H),3.07-3.01 (m, 2H), 2.80-2.76 (m, 1H), 2.74-2.64 (m, 2H), 2.64-2.58 (m,1H), 1.94 (s, 3H), 1.92-1.84 (m, 2H); ¹³C NMR (100 MHz, CD₃OD) δ 173.65,143.71, 129.57, 129.46, 126.88, 81.26, 81.08, 70.48, 69.44, 60.82,47.59, 39.59, 34.70, 31.25, 22.81. MS, (ES, m/z) [M+H]⁺ 323.21.

Example 2N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-(4-methoxyphenyl)propyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-(4-methoxyphenyl)propyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, using3-(4-methoxyphenyl)propionaldehyde. Colorless oil. Yield 48%. ¹H NMR(500 MHz, CD₃OD) δ 7.13-7.11 (m, 2H), 6.82-6.80 (m, 2H), 3.98-3.97 (m,1H), 3.79-3.78 (m, 1H), 3.75 (s, 3H), 3.72 (dd, J=5.1, 11.3 Hz, 1H),3.65 (dd, J=3.3, 11.3 Hz, 1H), 3.48 (dd, J=3.2, 13.6 Hz, 1H), 3.15 (dd,J=7.0, 13.6 Hz, 1H), 3.05-3.00 (m, 2H), 2.78-2.73 (m, 1H), 2.70-2.62 (m,2H), 2.57-2.51 (m, 1H), 1.95 (s, 3H), 1.88-1.79 (m, 2H); ¹³C NMR (125MHz, CD₃OD) δ 173.66, 159.39, 135.61, 130.46, 114.82, 81.23, 81.04,70.39, 69.34, 60.74, 55.72, 47.51, 39.48, 33.75, 31.36, 22.80. MS, (ES,m/z) [M+H]⁺ 353.21.

Example 3N-(((2R,3R,4R,5R)-1-(3-([1,1′-biphenyl]-4-yl)propyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

To a solution of [1,1′-biphenyl]-4-carbaldehyde (1.82 g, 10.0 mmol) and(carbomethoxymethyl)-triphenylphosphonium bromide (8.30 g, 20.0 mmol) inDCM (100 mL) and water (80 mL) was added NaOH (2 N, 10 mL, 20 mmol).After the mixture was stirred at room temperature for 3 h, the organiclayer was collected and the aqueous was extracted with DCM (2×30 mL).The combined extract was dried over anhydrous Na₂SO₄. After filtrationthe solvent was evaporated under reduced pressure, and the residue waspurified on silica gel by automatic flash column chromatography(DCM/hexanes, 1:2 to 2:1) to afford (E)-methyl3-([1,1′-biphenyl]-4-yl)acrylate as a white solid (2.00 g, 84%). ¹H NMR(500 MHz, CDCl₃) δ 7.74 (d, J=16.0 Hz, 1H), 7.64-7.59 (m, 6H), 7.48-7.44(m, 2H), 7.40-7.36 (m, 1H), 6.48 (d, J=16.0 Hz, 1H), 3.83 (s, 3H).

A suspension of (E)-methyl 3-([1,1′-biphenyl]-4-yl)acrylate (2.00 g,8.39 mmol) and Pd(OH)₂/C (20% in Pd, 0.40 g, 0.75 mmol) in dry methanol(100 mL) was stirred under an atmosphere of hydrogen for 3 h. Thesuspension was then filtered through a celite cake, and the filtrate wasconcentrated under reduced pressure to give pure methyl3-([1,1′-biphenyl]-4-yl)propanoate as a white solid (2.01 g, 100%). ¹HNMR (500 MHz, CDCl₃) δ 7.61-7.59 (m, 2H), 7.56-7.54 (m, 2H), 7.47-7.44(m, 2H), 7.38-7.36 (m, 1H), 7.31-7.29 (m, 2H), 3.72 (s, 3H), 3.03 (t,J=7.8 Hz, 2H), 2.71 (t, J=7.8 Hz, 2H).

At 0° C., under argon, to solution of methyl3-([1,1′-biphenyl]-4-yl)propanoate (2.00 g, 8.33 mmol) in dry THF (100mL) was added lithium aluminum hydride (LAH) (0.38 g, 10 mmol). Afterthe mixture was stirred at 0° C. for 2 h NaOH (5 N, 2 mL) was added toquench the reaction and the resulted suspension was stirred at roomtemperature for 1 h. The suspension was dried over anhydrous Na₂SO₄,then filtered through a celite cake. The filtrate was concentrated underreduced pressure to give pure 3-([1,1′-biphenyl]-4-yl)propan-1-ol as awhite solid (1.76 g, 100%). ¹H NMR (500 MHz, CDCl₃) δ 7.60-7.58 (m, 2H),7.54-7.52 (m, 2H), 7.44-7.41 (m, 2H), 7.35-7.31 (m, 1H), 7.29-7.27 (m,2H), 3.72 (t, J=6.4 Hz, 2H), 2.76 (t, J=7.7 Hz, 2H), 1.97-1.91 (m, 2H).

A suspension of 3-([1,1′-biphenyl]-4-yl)propan-1-ol (1.76 g, 8.30 mmol)and Dess-Martin periodinane (5.30 g, 12.5 mmol) in DCM (80 mL) wasstirred at room temperature for 1 h. Diethyl ether (200 mL) was added,and the resulted white solid was filtered off through a celite cake. Theorganic filtrate was washed with mixed saturated NaHCO₃ and Na₂S₂O₃(40/5 mL), and then was dried over anhydrous Na₂SO₄. After filtrationthe solvent was evaporated under reduced pressure, and the residue waspurified on silica gel by automatic flash column chromatography(EtOAc/hexanes, 1:9 to 1:5) to afford 3-([1,1′-biphenyl]-4-yl)propanalas a pale yellow solid (1.40 g, 80%). ¹H NMR (500 MHz, CDCl₃) δ 9.86 (t,J=1.2 Hz, 1H), 7.74-7.56 (m, 2H), 7.54-7.52 (m, 2H), 7.45-7.41 (m, 2H),7.35-7.32 (m, 1H), 7.28-7.26 (m, 2H), 3.01 (t, J=7.6 Hz, 2H), 2.82 (dt,J=1.2, 7.6 Hz, 2H).

N-(((2R,3R,4R,5R)-1-(3-([1,1-biphenyl]-4-yl)propyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, using3-([1,1′-biphenyl]-4-yl)propanal. Off-white solid. Yield 84%. ¹H NMR(500 MHz, CD₃OD) δ 7.59-7.56 (m, 2H), 7.53-7.51 (m, 2H), 7.42-7.39 (m,2H), 7.31-7.28 (m, 3H), 3.99-3.97 (m, 1H), 3.80-3.78 (m, 1H), 3.75 (dd,J=5.1, 11.3 Hz, 1H), 3.67 (dd, J=3.3, 11.3 Hz, 1H), 3.50 (dd, J=3.8,13.7 Hz, 1H), 3.18 (dd, J=6.5, 13.7 Hz, 1H), 3.08-3.05 (m, 2H),2.85-2.63 (m, 4H), 1.95 (s, 3H), 1.95-1.86 (m, 2H); ¹³C NMR (125 MHz,CDCl₃+CD₃OD) δ 172.32, 141.00, 140.98, 138.93, 128.79, 128.78, 127.14,127.12, 126.97, 79.67, 79.32, 69.22, 68.68, 59.68, 46.50, 38.63, 33.11,29.80, 22.70. MS, (ES, m/z) [M+H]⁺ 399.23.

Example 4N-(((2R,3R,4R,5R)-1-(2-([1,1′-biphenyl]-4-yl)ethyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-(2-([1,1-biphenyl]-4-yl)ethyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, using2-([1,1′-biphenyl]-4-yl)acetaldehyde which was obtained using aliterature procedure (J. Med. Chem. 2009, 52(9), 2776-2785). Whitesolid. Yield 58%. ¹H NMR (500 MHz, CD₃OD) δ 7.60-7.57 (m, 2H), 7.56-7.53(m, 2H), 7.43-7.39 (m, 2H), 7.36-7.33 (m, 2H), 7.30-7.31 (m, 1H),3.99-3.98 (m, 1H), 3.87 (dd, J=4.6, 11.6 Hz, 1H), 3.76 (dd, J=3.6, 11.6Hz, 1H), 3.76-3.74 (m, 1H), 3.56-3.52 (m, 1H), 3.24-3.22 (m, 1H),3.17-3.11 (m, 3H), 2.98-2.92 (m, 2H), 2.87-2.83 (m, 1H), 1.80 (s, 3H);¹³C NMR (125 MHz, CD₃OD) δ 173.72, 142.37, 141.03, 140.46, 130.52,129.98, 128.29, 128.09, 127.92, 80.75, 80.72, 70.15, 69.69, 60.80,49.78, 39.29, 35.16, 22.82. MS, (ES, m/z) [M+H]⁺ 385.22.

Example 5N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(4-phenylbutyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(4-phenylbutyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, using4-phenylbutanal which was obtained using a literature procedure (J. Am.Chem. Soc. 1996, 118(3), 502-508). Pale yellow oil. Yield 72%. ¹H NMR(500 MHz, CD₃OD) δ 7.25-7.22 (m, 2H), 7.20-7.17 (m, 2H), 7.15-7.12 (m,1H), 3.98-3.97 (m, 1H), 3.80-3.79 (m, 1H), 3.76 (dd, J=4.9, 11.5 Hz,1H), 3.68 (dd, J=3.6, 11.5 Hz, 1H), 3.49 (dd, J=3.4, 13.5 Hz, 1H), 3.23(dd, J=6.7, 13.8 Hz, 1H), 3.10-3.05 (m, 2H), 2.85-2.82 (m, 1H),2.77-2.73 (m, 1H), 2.66-2.61 (m, 2H), 1.92 (s, 3H), 1.72-1.58 (m, 3H);¹³C NMR (125 MHz, CD₃OD) δ 173.81, 143.86, 129.55, 129.43, 126.85,81.00, 80.75, 70.53, 69.73, 60.62, 48.10, 39.53, 36.80, 30.50, 28.73,22.78. MS, (ES, m/z) [M+H]⁺ 337.21.

Example 6N-(((2R,3R,4R,5R)-1-(3,3-diphenylpropyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-(3,3-diphenylpropyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination, using3,3-diphenylpropanal. ¹H NMR (500 MHz, CD₃OD) δ 7.36-7.24 (m, 8H),7.20-7.17 (m, 2H), 4.13-4.07 (m, 1H), 4.02-3.98 (br, 1H), 3.84-3.80 (br,1H), 3.71-3.61 (m, 2H), 3.48-3.40 (m, 1H), 3.35-3.32 (m, 1H), 3.12-3.04(m, 3H), 2.81-2.74 (m, 1H), 2.71-2.63 (m, 1H), 2.50-2.40 (m, 1H),2.32-2.23 (m, 1H), 1.96 (s, 3H); ¹³C NMR (125 MHz, CD₃OD) δ 172.2,145.3, 144.5, 128.1, 128.0, 127.6, 127.4, 125.9, 125.7, 79.5, 79.4,69.2, 68.1, 59.2, 48.8, 45.2, 37.9, 33.5, 21.3; MS (ES, m/z) [M+H]⁺399.2.

Example 7N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(pyridin-3-ylmethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(pyridin-3-ylmethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination using3-pyridinecarboxaldehyde. Colorless oil. Yield 60%. ¹H NMR (500 MHz,CD₃OD) δ 8.57 (s, 1H), 8.40 (d, J=4.9 Hz, 1H), 7.92 (d, J=7.8 Hz, 1H),7.40 (dd, J=4.9, 7.8 Hz, 1H), 4.07-3.99 (m, 3H), 3.85-3.83 (m, 1H), 3.73(dd, J=4.6, 11.5 Hz, 1H), 3.66 (dd, J=3.4, 11.5 Hz, 1H), 3.44 (dd,J=3.4, 13.9 Hz, 1H), 3.33-3.29 (m, 1H), 3.10-3.07 (m, 1H), 2.99-2.97 (m,1H), 1.93 (s, 3H); ¹³C NMR (125 MHz, CD₃OD) δ 173.59, 150.05, 148.63,138.40, 137.87, 125.25, 81.18, 80.77, 70.02, 69.65, 61.36, 49.56, 40.28,22.80. MS, (ES, m/z) [M+H]⁺ 296.16.

Example 8N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(pyridin-2-ylmethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(pyridin-2-ylmethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination using2-pyridinecarboxaldehyde. Pale yellow oil. Yield 35%. ¹H NMR (500 MHz,CD₃OD) δ 8.50-8.49 (m, 1H), 7.82 (dt, J=1.7, 7.7 Hz, 1H), 7.57 (d, J=7.7Hz, 1H), 7.33-7.30 (m, 1H), 4.14-4.08 (m, 2H), 4.01 (t, J=3.2 Hz, 1H),3.81 (t, J=3.2 Hz, 1H), 3.74 (dd, J=4.4, 11.7 Hz, 1H), 3.64 (dd, J=3.2,11.7 Hz, 1H), 3.53 (dd, J=3.2, 14.0 Hz, 1H), 3.24 (dd, J=5.8, 14.0 Hz,1H), 3.11-3.09 (m, 1H), 3.04-3.02 (m, 1H), 1.94 (s, 3H). MS, (ES, m/z)[M+H]⁺ 296.16.

Example 9N-(((2R,3R,4R,5R)-1-([1,1′-biphenyl]-4-ylmethyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamide

N-(((2R,3R,4R,5R)-1-([1,1′-biphenyl]-4-ylmethyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)methyl)acetamidewas prepared using the general procedure for reductive amination using[1,1′-biphenyl]-4-carbaldehyde. Off-white solid. Yield 90%. ¹H NMR (500MHz, CD₃OD) δ 7.59-7.55 (m, 4H), 7.49-7.47 (m, 2H), 7.42-7.39 (m, 2H),7.32-7.29 (m, 1H), 4.04-4.03 (m, 1H), 4.01-3.95 (m, 2H), 3.85-3.84 (m,1H), 3.79 (dd, J=4.9, 11.4 Hz, 1H), 3.66 (dd, J=3.1, 11.4 Hz, 1H), 3.44(dd, J=3.4, 13.8 Hz, 1H), 3.32 (dd, J=6.1, 13.8 Hz, 1H), 3.15-3.13 (m,1H), 3.07-3.05 (m, 1H), 1.93 (s, 3H); ¹³C NMR (125 MHz, CD₃OD) δ 173.61,142.40, 141.28, 140.20, 129.95, 129.93, 128.32, 128.06, 127.99, 81.34,80.88, 70.06, 69.80, 61.23, 52.09, 40.38, 22.84. MS, (ES, m/z) [M+H]⁺371.20.

Example 102-((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide

At 0° C., to a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyrrole1-oxide (Tetrahedron 2009, 65, 93-100) (5.80 g, 13.9 mmol) in THF (60mL) was added allylmagnesium chloride (2 M, THF, 8.5 mL, 17 mmol). Afterthe mixture was stirred at room temperature for 16 h saturated aqueousNH₄Cl (50 mL) was added and the mixture was extracted with EtOAc (2×50mL). The combined filtrate was dried over anhydrous Na₂SO₄. Afterfiltration the solvent was evaporated under reduced pressure, and theresidue was purified on silica gel by automatic flash columnchromatography (EtOAc/hexanes, 1:9 to 1:3) to afford(2R,3R,4R,5R)-2-allyl-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)pyrrolidin-1-olas a pale yellow oil (4.80 g, 75%). ¹H NMR (500 MHz, CDCl₃) δ 7.37-7.28(m, 15H), 5.90-5.80 (m, 1H), 5.14-5.08 (m, 2H), 4.61-4.40 (m, 6H),3.99-3.97 (m, 1H), 3.88-3.86 (m, 1H), 3.81-3.74 (m, 1H), 3.69-3.65 (m,1H), 3.58-3.55 (m, 1H), 3.42-3.38 (m, 1H), 2.74-2.69 (m, 1H), 2.36-2.30(m, 1H). MS, (ES, m/z) [M+H]⁺ 460.2.

A suspension of(2R,3R,4R,5R)-2-allyl-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)pyrrolidin-1-ol(5.89 g, 12.8 mmol), Zn powder (2.20 g, 34.7 mmol) in AcOH/MeOH (9mL/100 mL) was stirred at reflux for 1 h. After being cooled to roomtemperature, the suspension was filtered thought a celite cake andconcentrated. The residue was treated with saturated aqueous NaHCO₃ (80mL), and the extracted with DCM (3×80 mL). The combined filtrate wasdried over anhydrous Na₂SO₄. After filtration the solvent was evaporatedunder reduced pressure, and the residue was dissolved in DCM (100 mL).To the solution was added Boc₂O (4.36 g, 20.0 mmol) and DIPEA (2.0 mL),and the mixture was stirred at room temperature for 16 h. Afterconcentration the residue was purified on silica gel by automatic flashcolumn chromatography (EtOAc/hexanes, 1:20 to 1:5) to afford(2R,3R,4R,5R)-tert-butyl2-allyl-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)pyrrolidine-1-carboxylateas a colorless oil (5.90 g, 85%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 544.3.

To a solution of (2R,3R,4R,5R)-tert-butyl2-allyl-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)pyrrolidine-1-carboxylate(5.70 g, 10.5 mmol) and 4-methyl morpholine-N-oxide (2.46 g, 21.0 mmol)in acetone/H₂O (60 mL/10 mL) was added OsO₄ (4% in water, 0.60 mL, 24mg, 0.094 mmol). After the mixture was stirred at room temperature for16 h a solution of Na₂S₂O₃ (5 g) in water (10 mL) was added. Afterstirring at room temperature for 20 min the mixture was concentrated andextracted with DCM (3×100 mL). The combined filtrate was dried overanhydrous Na₂SO₄. After filtration the solvent was evaporated underreduced pressure, and the residue was purified on silica gel byautomatic flash column chromatography (EtOAc/hexanes, 1:2 to 2:1) toafford (2R,3R,4R,5R)-tert-butyl3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2,3-dihydroxypropyl)pyrrolidine-1-carboxylateas a pale yellow oil (6.02 g, 99%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 578.3.

To a suspension of (2R,3R,4R,5R)-tert-butyl3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2,3-dihydroxypropyl)pyrrolidine-1-carboxylate(5.54 g, 10.2 mmol) in THF (60 mL) and an aqueous NaH₂PO₄/NaHCO₃ buffersolution (1:1 molar ratio, 60 mL) was added NaIO₄ (3.46 g, 16.2 mmol),and the mixture was stirred at room temperature for 3 h. The mixture wasthen diluted with brine (100 mL) and extracted with EtOAc (3×80 mL). Thecombined filtrate was dried over anhydrous Na₂SO₄. After filtration thesolvent was evaporated under reduced pressure, and the residue waspurified on silica gel by automatic flash column chromatography(EtOAc/hexanes, 1:10 to 1:3) to afford (2R,3R,4R,5R)-tert-butyl3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2-oxoethyl)pyrrolidine-1-carboxylateas a colorless oil (5.10 g, 92%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 546.3.

To a solution of (2R,3R,4R,5R)-tert-butyl3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2-oxoethyl)pyrrolidine-1-carboxylate(5.01 g, 9.18 mmol), 2-methylbut-2-ene (3.2 g, 46 mmol) in mixedtert-butanol/CDCl₃ (12 mL/75 mL) was added a solution of NaH₂PO₄ (4.0 g,33 mmol) in water (7 mL) and NaClO₂ (1.56 g, 17.2 mmol). After themixture was stirred at room temperature for 3 h a solution of Na₂S₂O₃ (5g) in water (10 mL) was added. After stirring at room temperature for 10min the mixture was adjusted to weakly acidic (pH=˜3) with 1 N HCl, andthen extracted with DCM (3×50 mL). The combined filtrate was dried overanhydrous Na₂SO₄. After filtration the solvent was evaporated underreduced pressure, and the residue was purified on silica gel byautomatic flash column chromatography (EtOAc/hexanes, 1:4 to 1:2) toafford2-((2R,3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)aceticacid as a colorless oil (5.02 g, 97%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 562.3.

A mixture of2-((2R,3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)aceticacid (2.10 g, 3.74 mmol), MeNH₂ hydrochloride (0.54 g, 8.0 mmol), DIPEA(2.3 g, 18 mmol) and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate methanaminium (HATU) (1.90 g, 5.00 mmol) inDMF (20 mL) was stirred at room temperature for 2 days. The mixture wasdiluted with brine (100 mL) and extracted with Et₂O (3×50 mL). Thecombined filtrate was washed with saturated aqueous NaHCO₃ (50 mL), anddried over anhydrous Na₂SO₄. After filtration the solvent was evaporatedunder reduced pressure, and the residue was purified on silica gel byautomatic flash column chromatography (EtOAc/hexanes, 1:2 to 1:1) toafford (2R,3R,4R,5R)-tert-butyl3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2-(methylamino)-2-oxoethyl)pyrrolidine-1-carboxylateas a colorless oil (2.15 g, 100%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 575.3.

To a solution of (2R,3R,4R,5R)-tert-butyl3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2-(methylamino)-2-oxoethyl)pyrrolidine-1-carboxylate(2.15 g, 3.74 mmol) in mixed methanol/EtOAc (40 mL/20 mL) was bubbledwith HCl gas for 30 seconds. After stirring for 2 h the mixture wasconcentrated. The residue was treated with saturated aqueous NaHCO₃ (40mL), and extracted with DCM (3×50 mL). The combined filtrate was driedover anhydrous Na₂SO₄. After filtration the solvent was evaporated underreduced pressure to afford pure2-((2R,3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)pyrrolidin-2-yl)-N-methylacetamideas a colorless oil (1.70 g, 96%). ¹H NMR (500 MHz, CDCl₃) δ 7.34-7.25(m, 15H), 4.52-4.50 (m, 6H), 3.91-3.89 (m, 1H), 3.78-3.76 (m, 1H),3.56-3.53 (m, 3H), 3.48 (s, 1H), 3.41-3.38 (m, 1H), 2.74 (d, J=4.8 Hz,3H), 2.51 (dd, J=9.4, 15.7, 1H), 2.40 (dd, J=4.3, 15.7, 1H.

To a solution of2-((2R,3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)pyrrolidin-2-yl)-N-methylacetamide(1.40 g, 2.95 mmol) in DCM (100 mL) at −78° C. was added BCl₃ (1.0 M,17.7 mL, 17.7 mmol), and the solution was stirred at room temperaturefor 16 h. After cooling at −78° C. the solution was quenched with MeOH.After concentration under reduced pressure the residue(2-((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N-methylacetamide)as an impure HCl salt (0.87 g) was obtained. The residue (0.30 g) wasdissolved in MeOH (10 mL), and to which hydrocinnamaldehyde (0.334 g,2.50 mmol), NaBH₃CN (0.20 g, 3.0 mmol) was added. The mixture wasstirred at room temperature for 16 h, and then concentrated underreduced pressure. The residue was purified on silica gel by flash columnchromatography (EtOAc; then MeOH/DCM, 1:5) to afford2-((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamideas a white foam (0.310 g, 94%). ¹H NMR (500 MHz, CD₃OD) δ 7.26-7.23 (m,2H), 7.22-7.20 (m, 2H), 7.16-7.13 (m, 1H), 3.96-3.95 (m, 1H), 3.80-3.79(m, 1H), 3.71-3.65 (m, 2H), 3.41-3.39 (m, 1H), 2.89-2.87 (m, 1H),2.73-2.65 (m, 6H), 2.61-2.56 (m, 1H), 2.47 (dd, J=4.9, 14.7 Hz, 1H),2.36 (dd, J=8.2, 14.7 Hz, 1H), 1.88-1.80 (m, 2H); ¹³C NMR (125 MHz,CD₃OD) δ 175.06, 143.59, 129.58, 129.46, 126.92, 82.07, 81.10, 71.05,66.81, 61.07, 47.77, 35.12, 34.71, 30.97, 26.45. MS, (ES, m/z) [M+H]⁺323.2.

Example 112-((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-(4-methoxyphenyl)propyl)pyrrolidin-2-yl)-N-ethylacetamide

2-((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N-ethylacetamidewas prepared from2-((2R,3R,4R,5R)-3,4-bis(benzyloxy)-5-((benzyloxy)methyl)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)aceticacid using procedures analagous to those described for Example 10.2-((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N-ethylacetamide(50 mg, 0.2 mmol) was dissolved in MeOH (3 mL), to which3-(4-methoxyphenyl)propionaldehyde (65 mg, 0.40 mmol) and NaBH₃CN (25mg, 0.40 mmol) were added. The mixture was stirred at room temperaturefor 16 h, and then concentrated under reduced pressure. The residue waspurified on silica gel by flash column chromatography to afford productas white foam (36 mg, 55%). ¹H NMR (500 MHz, CD₃OD) δ 7.13 (d, J=9.0 Hz,2H), 6.83 (d, J=9.0 Hz, 2H), 3.99-3.96 (br, 1H), 3.84-3.81 (br, 1H),3.77 (s, 3H), 3.73-3.66 (m, 2H), 3.44-3.39 (m, 1H), 3.21 (q, J=7.0 Hz,2H), 2.92-2.87 (m, 1H), 2.75-2.62 (m, 3H), 2.60-2.52 (m, 1H), 2.51-2.45(m, 1H), 2.40-2.33 (m, 1H), 1.90-1.75 (m, 2H)), 1.13 (t, J=7.2 Hz, 3H);MS (ES, m/z) [M+H]⁺ 367.2.

Example 122-((2R,3R,4R,5R)-1-(3,3-diphenylpropyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N-ethylacetamide

2-((2R,3R,4R,5R)-1-(3,3-diphenylpropyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N-ethylacetamidewas prepared from2-((2R,3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N-ethylacetamideand 3,3-diphenylpropanal using procedures analogous to those describedfor Example 11. ¹H NMR (500 MHz, CD₃OD) δ 7.35-7.24 (m, 8H), 7.18-7.13(m, 2H), 4.08-4.03 (m, 1H), 3.99-3.96 (br, 1H), 3.84-3.78 (br, 1H),3.63-3.55 (m, 2H), 3.47-3.42 (m, 1H), 3.35-3.32 (m, 1H), 3.24-3.12 (m,2H), 2.86-2.82 (m, 1H), 2.65-2.58 (m, 2H), 2.43-2.33 (m, 2H), 2.32-2.20(m, 2H), 1.11 (t, J=7.2 Hz, 3H); ¹³C NMR (125 MHz, CD₃OD) δ 172.7,145.2, 144.7, 128.1, 128.0, 127.7, 127.4, 125.8, 125.7, 80.5, 79.6,69.6, 65.1, 59.6, 48.9, 45.1, 33.8, 33.7, 33.5, 13.4; MS (ES, m/z)[M+H]⁺ 413.2.

Example 132-((2R,3S,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide

A suspension of (2R,3R,4R,5R)-tert-butyl3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2-(methylamino)-2-oxoethyl)pyrrolidine-1-carboxylate(1.35 g, 2.35 mmol) and Pd on carbon (10% Pd by weight, 0.320 g, 0.301mmol) in MeOH (25 mL) was shaken at room temperature under hydrogen (40psi) for 16 h. The suspension was then filtered through a celite cake,and the filtrate was concentrated to afford pure(2R,3R,4R,5R)-tert-butyl3,4-dihydroxy-2-(hydroxymethyl)-5-(2-(methylamino)-2-oxoethyl)pyrrolidine-1-carboxylateas a white solid (0.72 g, 100%). This compound exists as twoconformational rotamers in CD₃OD and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 305.2, [M+Na]⁺ 327.2.

To a solution of (2R,3R,4R,5R)-tert-butyl3,4-dihydroxy-2-(hydroxymethyl)-5-(2-(methylamino)-2-oxoethyl)pyrrolidine-1-carboxylate(0.300 g, 0.982 mmol) and imidazole (0.20 g, 3.0 mmol) in anhydrouspyridine (15 mL), at 0° C., was added1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane mixture (TIPDSCl) (0.370g, 1.17 mmol), and the mixture was stirred at room temperature for 5 h.The mixture was diluted with brine (50 mL) and extracted with EtOAc(3×30 mL). The combined filtrate was dried over anhydrous Na₂SO₄. Afterfiltration the solvent was evaporated under reduced pressure, and theresidue was purified on silica gel by automatic flash columnchromatography (EtOAc/hexanes, 2:3 to 3:2) to afford(6aR,8R,9R,9aR)-tert-butyl9-hydroxy-2,2,4,4-tetraisopropyl-8-(2-(methylamino)-2-oxoethyl)tetrahydro-[1,3,5,2,4]trioxadisilocino[7,6-b]pyrrole-7(8H)-carboxylateas a white solid (0.44 g, 82%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 547.3.

A mixture of (6aR,8R,9R,9aR)-tert-butyl9-hydroxy-2,2,4,4-tetraisopropyl-8-(2-(methylamino)-2-oxoethyl)tetrahydro-[1,3,5,2,4]trioxadisilocino[7,6-b]pyrrole-7(8H)-carboxylate(0.440 g, 0.805 mmol) and Dess-Martin periodinane (DMP) (0.636 g, 1.50mmol) in DCM (60 mL) was stirred at room temperature for 1 h. Themixture was diluted with Et₂O (60 mL) and filtered through a celitecake. The filtrate was concentrated and treated with saturated aqueousNaHCO₃ (20 mL). The mixture was extracted with EtOAc (2×20 mL). Thecombined filtrate was dried over anhydrous Na₂SO₄. After filtration thesolvent was evaporated under reduced pressure, and the residue waspurified on silica gel by automatic flash column chromatography(EtOAc/hexanes, 1:2 to 1:1) to afford (6aR,8R,9aR)-tert-butyl2,2,4,4-tetraisopropyl-8-(2-(methylamino)-2-oxoethyl)-9-oxotetrahydro-[1,3,5,2,4]trioxadisilocino[7,6-b]pyrrole-7(8H)-carboxylateas a white solid (0.35 g, 80%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 545.3.

To a solution of (6aR,8R,9aR)-tert-butyl2,2,4,4-tetraisopropyl-8-(2-(methylamino)-2-oxoethyl)-9-oxotetrahydro-[1,3,5,2,4]trioxadisilocino[7,6-b]pyrrole-7(8H)-carboxylate(0.830 g, 1.52 mmol), at 0° C., was added NaBH₄ (0.114 g, 3.00 mmol),and the mixture was stirred at 0° C. for 20 min. The mixture wasquenched with water and concentrated. The residue was diluted withsaturated aqueous NaHCO₃ (20 mL) and extracted with DCM (3×40 mL). Thecombined filtrate was dried over anhydrous Na₂SO₄. After filtration thesolvent was evaporated under reduced pressure, and the residue waspurified on silica gel by automatic flash column chromatography(EtOAc/hexanes, 2:3 to 3:2) to afford (6aR,8R,9S,9aR)-tert-butyl9-hydroxy-2,2,4,4-tetraisopropyl-8-(2-(methylamino)-2-oxoethyl)tetrahydro-[1,3,5,2,4]trioxadisilocino[7,6-b]pyrrole-7(8H)-carboxylateas a white solid (0.80 g, 95%). This compound exists as twoconformational rotamers in CDCl₃ and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 547.3.

To a solution of (6aR,8R,9S,9aR)-tert-butyl9-hydroxy-2,2,4,4-tetraisopropyl-8-(2-(methylamino)-2-oxoethyl)tetrahydro-[1,3,5,2,4]trioxadisilocino[7,6-b]pyrrole-7(8H)-carboxylate(0.590 g, 1.08 mmol), at 0° C., was added TBAF (1.0 M, 3.5 mL, 3.5mmol), and the mixture was stirred at room temperature for 3 h. Afterconcentration under reduced pressure the residue was purified on silicagel by automatic flash column chromatography (MeOH/EtOAc, 1:20 to 1:10)to afford (2R,3R,4S,5R)-tert-butyl3,4-dihydroxy-2-(hydroxymethyl)-5-(2-(methylamino)-2-oxoethyl)pyrrolidine-1-carboxylateas a white solid (0.300 g, 91%). This compound exists as twoconformational rotamers in CD₃OD and gives a complicated ¹H NMR pattern.MS, (ES, m/z) [M+H]⁺ 306.2.

To a solution of (2R,3R,4S,5R)-tert-butyl3,4-dihydroxy-2-(hydroxymethyl)-5-(2-(methylamino)-2-oxoethyl)pyrrolidine-1-carboxylate(0.190 g, 0.623 mmol) in methanol (10 mL) was bubbled with HCl gas for30 seconds. After stirring for 2 h the mixture was concentrated andwashed with DCM. The residue was dissolved in MeOH (3 mL), andhydrocinnamaldehyde (0.134 g, 1.00 mmol), NaBH₃CN (0.10 g, 1.5 mmol)were added to the resulting solution. The mixture was stirred at roomtemperature for 16 h, and then concentrated under reduced pressure. Theresidue was treated with saturated HCl in methanol for 30 min, and thenconcentrated to dryness. The residue was then neutralized with excessDIPEA and methylamine HCl salt in isopropanol, and heated at reflux for16 h. After concentration the residue was purified on silica gel byflash column chromatography (EtOAc; then MeOH/DCM, 1:6) to afford2-((2R,3S,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamideas an off-white foam (0071 g, 36%). ¹H NMR (500 MHz, CD₃OD) δ 7.30-7.27(m, 2H), 7.24-7.22 (m, 2H), 7.20-7.17 (m, 1H), 4.23-4.19 (m, 2H),4.00-3.98 (m, 1H), 3.88-3.80 (m, 2H), 3.53-3.51 (m, 1H), 3.29-3.28 (m,1H), 3.22-3.18 (m, 1H), 2.89-2.84 (m, 1H), 2.76-2.67 (m, 6H), 2.11-2.09(m, 1H), 2.02-1.97 (m, 1H); ¹³C NMR (125 MHz, CD₃OD) δ 172.48, 141.95,129.74, 129.57, 127.47, 73.07, 72.45, 72.25, 66.31, 59.70, 52.60, 33.87,31.86, 28.88, 26.58. MS, (ES, m/z) [M+H]⁺ 323.2.

The following examples are synthesized according to procedures analogousto the schemes and examples outlined herein.

TABLE 3 Example Name Structure 14 2-((2R,3R,4R,5R)-1-(3-([1,1′-biphenyl]-4-yl)propyl)-3,4-dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- ethylacetamide

15 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-(4′-sulfamoyl-[1,1′-biphenyl]-4-yl)propyl)pyrrolidin- 2-yl)-N-methylacetamide

16 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-phenethylpyrrolidin-2-yl)-N- ethylacetamide

17 (2S,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-phenethyl-N-propylpyrrolidine-2-carboxamide

18 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-phenethylpyrrolidin-2- yl)methyl)propionamide

19 N-ethyl-2-((2R,3R,4R,5S)-5- (fluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin-2-yl)acetamide

20 N-ethyl-2-((2R,3R,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-phenethylpyrrolidin-2-yl)acetamide

21 N-ethyl-2-((2R,3S,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-phenethylpyrrolidin-2-yl)acetamide

22 2-((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin- 2-yl)-N-ethylacetamide

23 2-((2R,3R,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

24 2-((2R,3R,4R,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

25 2-((2R,3S,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

26 2-((2R,3R,4S,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

27 2-((2R,3R,4R,5S)-5-(fluoromethyl)- 3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

28 2-((2R,3R,4R,5S)-5-(difluoromethyl)- 3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

29 2-((2R,3R,4R,5R)-3,4-dihydroxy-5-methyl-1-(3-phenylpropyl)pyrrolidin- 2-yl)-N-methylacetamide

30 2-((2S,4S,5R)-4-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

31 2-((2R,3S,5S)-3-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

32 2-((2R,3R,4R)-3,4-dihydroxy-1-(3- phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide

33 2-((2R,3R,4R,5S)-3-fluoro-5- (fluoromethyl)-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

34 2-((2R,3S,4R,5S)-5-(difluoromethyl)- 3-fluoro-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N- methylacetamide

35 N-(((2R,3R,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

36 N-(((2R,3R,4R,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

37 N-(((2R,3S,4R,5R)-3-fluoro-4- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

38 N-(((2R,3R,4S,5R)-4-fluoro-3- hydroxy-5-(hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

39 N-(((2R,3R,4R,5S)-5-(fluoromethyl)- 3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

40 N-(((2R,3R,4R,5S)-5- (difluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

41 N-(((2S,4S,5R)-4-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

42 N-(((2R,3S,5S)-3-hydroxy-5- (hydroxymethyl)-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

43 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-methyl-1-(3-phenylpropyl)pyrrolidin- 2-yl)methyl)propionamide

44 N-(((2R,3R,4R)-3,4-dihydroxy-1-(3- phenylpropyl)pyrrolidin-2-yl)methyl)propionamide

45 N-(((2R,3R,4R,5S)-3-fluoro-5- (fluoromethyl)-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

46 N-(((2R,3S,4R,5S)-5- (difluoromethyl)-3-fluoro-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2- yl)methyl)propionamide

47 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-phenylpropanoyl)pyrrolidin-2- yl)methyl)propionamide

48 N-(((2R,3R,4R,5R)-1-(3-(6- fluoropyridin-3-yl)propyl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2- yl)methyl)propionamide

49 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-2- yl)methyl)propionamide

50 N-(((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-(5-(trifluoromethoxy)benzo[d]thiazol-2- yl)propyl)pyrrolidin-2-yl)methyl)propionamide

51 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-((Z)-5,5,5-trifluoropent-3-en-1-yl)pyrrolidin-2- yl)-N-methylacetamide

52 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(pent-4-yn-1-yl)pyrrolidin-2-yl)-N- methylacetamide

53 2-((2R,3R,4R,5R)-1-butyryl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

54 2-((2R,3R,4R,5R)-1-(3-(6- fluoropyridin-3-yl)propanoyl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

55 2-((2R,3R,4R,5R)-1-cinnamyl-3,4- dihydroxy-5-(hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

56 2-((2R,3R,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)-1-(3-phenylprop-2-yn-1-yl)pyrrolidin-2-yl)-N- methylacetamide

57 2-((2R,3R,4R,5R)-1-((E)-3-(6- fluoropyridin-3-yl)allyl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

58 2-((2R,3R,4R,5R)-1-(3-(6- fluoropyridin-3-yl)prop-2-yn-1-yl)-3,4-dihydroxy-5- (hydroxymethyl)pyrrolidin-2-yl)-N- methylacetamide

Biological Activity

Assay for Determination of K_(I) Values for Inhibition of O-GlcNAcaseActivity

Experimental Procedure for Kinetic Analyses:

Enzymatic reactions were carried out in a reaction containing 50 mMNaH₂PO₄, 100 mM NaCl and 0.1% BSA (pH 7.0) using 2 mM4-Methylumbelliferyl N-acetyl-β-D-glucosaminide dihydrate (Sigma M2133)dissolved in ddH₂O, as a substrate. The amount of purified humanO-GlcNAcase enzyme used in the reaction was 0.7 nM. Test compound ofvarying concentrations was added to the enzyme prior to initiation ofthe reaction. The reaction was performed at room temperature in a96-well plate and was initiated with the addition of substrate. Theproduction of fluorescent product was measured every 60 sec for 45 minwith a Tecan Infinite M200 plate-reader with excitation at 355 nM andemission detected at 460 nM, with 4-Methylumbelliferone (Sigma M1381)used to produce a standard curve. The slope of product production wasdetermined for each concentration of compound tested and plotted, usingstandard curve fitting algorithms for sigmoidal dose response curves.The values for a four parameter logistic curve fit of the data weredetermined.

K_(I) values were determined using the Cheng-Prusoff equation; the K_(m)of O-GlcNAcase for substrate was 0.2 mM.

Many compounds of the invention exhibit K_(I) values for inhibition ofO-GlcNAcase in the range 0.1 nM-50 μM.

Assay for Determination of K_(I) Values for Inhibition ofβ-Hexosaminidase Activity

Experimental Procedure for Kinetic Analyses:

Enzymatic reactions were carried out in a reaction containing 50 mMNaH₂PO₄, 100 mM NaCl and 0.1% BSA (pH 7.0) using 2 mM4-Methylumbelliferyl N-acetyl-β-D-glucosaminide dihydrate (Sigma M2133)dissolved in ddH₂O, as a substrate. The amount of purified humanβ-hexosaminidase enzyme used in the reaction was 24 nM. Test compound ofvarying concentrations was added to the enzyme prior to initiation ofthe reaction. The reaction was performed at room temperature in a96-well plate and was initiated with the addition of substrate. Theproduction of fluorescent product was measured every 60 sec for 45 minwith a Tecan Infinite M200 plate-reader with excitation at 355 nM andemission detected at 460 nM, with 4-Methylumbelliferone (Sigma M1381)used to produce a standard curve. The slope of product production wasdetermined for each concentration of compound tested and plotted, usingstandard curve fitting algorithms for sigmoidal dose response curves.The values for a four parameter logistic curve fit of the data weredetermined.

K_(I) values were determined using the Cheng-Prusoff equation.

When tested in this assay, many of the compounds described hereinexhibit K_(I) values for inhibition of β-hexosaminidase in the range 10nM to greater than 100 uM.

The selectivity ratio for inhibition of O-GlcNAcase overβ-hexosaminidase is defined here as:K_(I)(β-hexosaminidase)/K_(I)(O-GlcNAcase)

In general, many of the compounds described herein exhibit a selectivityratio in the range of about 10 to 100000. Thus, many compounds of theinvention exhibit high selectivity for inhibition of O-GlcNAcase overβ-hexosaminidase.

Assay for Determination of Cellular Activity for Compounds that InhibitO-GlcNAcase Activity

Inhibition of O-GlcNAcase, which removes O-GlcNAc from cellularproteins, results in an increase in the level of O-GlcNAcylated proteinin cells. An increase in O-GlcNAcylated protein can be measured by anantibody, such as RL-2, that binds to O-GlcNAcylated protein. The amountof O-GlcNAcylated protein:RL2 antibody interaction can be measured byenzyme linked immunosorbant assay (ELISA) procedures.

A variety of tissue culture cell lines, expressing endogenous levels ofO-GlcNAcase, can be utilized; examples include rat PC-12, and humanU-87, or SK—N—SH cells. In this assay, rat PC-12 cells are plated in96-well plates with approximately 10,000 cells/well. Compounds to betested are dissolved in DMSO, either 2 or 10 mM stock solution, and thendiluted with DMSO and water in a two-step process using a Tecanworkstation. Cells are treated with diluted compounds for 24 h (5.4 μLinto 200 μL 1 well volume) to reach a final concentration of inhibitordesired to measure a compound concentration dependent response,typically, ten 3 fold dilution steps, starting at 10 μM are used todetermine a concentration response curve. To prepare a cell lysate, themedia from compound treated cells is removed, the cells are washed oncewith phosphate buffered saline (PBS) and then lysed for 5 minutes atroom temperature in 50 μL of Phosphosafe reagent (Novagen Inc, Madison,Wis.) with protease inhibitors and PMSF. The cell lysate is collectedand transferred to a new plate, which is then either coated to assayplates directly or frozen −80° C. until used in the ELISA procedure. Ifdesired, the total protein concentration of samples is determined using20 μL of the sample using the BCA method.

The ELISA portion of the assay is performed in a black Maxisorp 96-wellplate that is coated overnight at 4° C. with 100 μL/well of the celllysate (1:10 dilution of the lysate with PBS containing proteaseinhibitors, phosphatase inhibitors, and PMSF). The following day thewells are washed 3 times with 300 μL/well of Wash buffer (Tris-bufferedsaline with 0.1% Tween 20). The wells are blocked with 100 μL/wellBlocking buffer (Tris buffered saline w/0.05% Tween 20 and 2.5% Bovineserum albumin). Each well is then washed two times with 300 μL/well ofwash buffer. The anti O-GlcNAc antibody RL-2 (Abcam, Cambridge, Mass.),diluted 1:1000 in blocking buffer, is added at 100 μL/well. The plate issealed and incubated at 37° C. for 2 h with gentle shaking. The wellsare then washed 3-times with 300 μL/well wash buffer. To detect theamount of RL-2 bound horse-radish peroxidase (HRP) conjugated goatanti-mouse secondary antibody (diluted 1:3000 in blocking buffer) isadded at 100 μL/well. The plate is incubated for 60 min at 37° C. withgentle shaking. Each well is then washed 3-times with 300 μL/well washbuffer. The detection reagent is added, 100 μL/well of Amplex Ultra REDreagent (prepared by adding 30 μL of 10 mM Amplex Ultra Red stocksolution to 10 mL PBS with 18 μL 3% hydrogen peroxide, H₂O₂). Thedetection reaction is incubated for 15 minutes at room temperature andthen read with excitation at 530 nm and emission at 590 nm.

The amount of O-GlcNAcylated protein, as detected by the ELISA assay, isplotted for each concentration of test compound using standard curvefitting algorithms for sigmoidal dose response curves. The values for afour parameter logistic curve fit of the data are determined, with theinflection point of the curve being the potency value for the testcompound.

Assay for Determination of Apparent Permeability (P_(app))

Bi-directional transport is evaluated in LLC-PK1 cells in order todetermine apparent permeability (P_(app)). LLC-PK1 cells can form atight monolayer and therefore can be used to assess vectorial transportof compounds from basolateral to apical (B→A) and from apical tobasolateral (A→B).

To determine P_(app), LLC-PK1 cells are cultured in 96-well transwellculture plates (Millipore). Solutions containing the test compounds (1μM) are prepared in Hank's Balanced Salt Solution with 10 mM HEPES.Substrate solution (150 μL) is added to either the apical (A) or thebasolateral (B) compartment of the culture plate, and buffer (150 μL) isadded to the compartment opposite to that containing the compound. Att=3 h, 50 μL samples are removed from both sides of monolayers dosedwith test compound and placed in 96 well plates, scintillant (200 μL) orinternal standard (100 μL labetolol 1 μM) is added to the samples andconcentration is determined by liquid scintillation counting in aMicroBeta Wallac Trilux scintillation counter (Perkin Elmer LifeSciences, Boston, Mass.) or by LCMS/MS (Applied Biosystems SCIEX API5000 triple quadruple mass spectrometer). [³H]Verapamil (1 μM) is usedas the positive control. The experiment is performed in triplicate.

The apparent permeability, P_(app), is calculated by the followingformula for samples taken at t=3 h:

$P_{app} = {\frac{{Volume}\mspace{14mu}{of}\mspace{14mu}{Receptor}\mspace{14mu}{Chamber}\mspace{14mu}({mL})}{\left\lbrack {{Area}\mspace{14mu}{of}\mspace{14mu}{membrane}\mspace{14mu}\left( {cm}^{2} \right)} \right\rbrack\left\lbrack {{Initial}\mspace{14mu}{Concentration}\mspace{14mu}\left( {\mu\; M} \right)} \right\rbrack} \times \frac{\Delta\mspace{14mu}{in}\mspace{14mu}{Concentration}\mspace{14mu}\left( {\mu\; M} \right)}{\Delta\mspace{14mu}{in}\mspace{14mu}{Time}\mspace{20mu}(s)}}$Where: Volume of Receptor Chamber is 0.15 mL; Area of membrane is 0.11cm²; the Initial Concentration is the sum of the concentration measuredin the donor plus concentration measured in receiver compartments at t=3h; A in Concentration is concentration in the receiver compartment at 3h; and A in Time is the incubation time (3×60×60=10800 s). P_(app) isexpressed as 10⁻⁶ cm/s. The P_(app) (LLC-PK1 cells) are the average ofthe P_(app) for transport from A to B and P_(app) for transport from Bto A at t=3 h:

${P_{app}\left( {{LLC} - {{PK}\; 1\;{Cells}}} \right)} = \frac{{P_{app}\left( A\rightarrow B \right)} + {P_{app}\left( B\rightarrow A \right)}}{2}$

Representative data from the binding, cell-based, and permeabilityassays described above are shown in the following table. Certaincompounds of the invention exhibit superior potency or permeability inone or more of these assays.

TABLE 4 Cell-based Fluorescence- Papp LLC- ELISA based hOGA PK1 cellsExample Structure EC₅₀ (nM) Ki (nM) (10⁻⁶ cm/s) —

ND 108 ND —

ND 2020 ND —

ND 1030 ND —

ND 619 <1.6 —

ND 343 ND 1

ND 369 ND 2

ND 106 2.6 3

665 16 5.0 4

ND 79 4.4 5

ND 172 <1.6 6

ND 41 ND 7

ND 2690 ND 8

ND 3020 ND 9

ND 4560 ND 10

ND 55 ND 11

ND 132 ND 12

ND 60 ND 13

ND 8.3 ND

The present invention has been described with regard to one or moreembodiments. However, it will be apparent to persons skilled in the artthat a number of variations and modifications can be made withoutdeparting from the scope of the invention as defined in the claims.

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What is claimed is:
 1. A compound of Formula (I) or a pharmaceuticallyacceptable salt thereof:

wherein R¹ is selected from the group consisting of: CH₂NR⁸R⁹, whereinR⁸ is H and R⁹ is C₁₋₆ acyl, and (CH₂)_(n)C(O)NR¹⁰ ₂, wherein n is 1,and one R¹⁰ is H and the other R¹⁰ is C₁₋₆ alkyl; R² is H and R³ is OH,or R² is H and R³ is H, or R² is H and R³ is F, or R² is F and R³ is H,or R² is F and R³ is F, or R² is OH and R³ is H; R⁴ is H and R⁵ is OH,or R⁴ is H and R⁵ is H, or R⁴ is H and R⁵ is F, or R⁴ is F and R⁵ is H,or R⁴ is F and R⁵ is F, or R⁴ is OH and R⁵ is H; R⁶ is selected from thegroup consisting of: CH₃, CH₂F, and CHF₂; and R⁷ is selected from thegroup consisting of: C₁₋₁₈ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₇₋₂₀arylalkyl, C₈₋₂₀ arylalkenyl, and C₈₋₂₀ arylalkynyl, each optionallysubstituted from one to four substituents with one of more of halo, OH,OCF₃, CN, SO₂NH₂, SO₂Me, C(O)NH₂, CH₂F, CHF₂, CF₃, CH₂CH₂F, CH₂CF₃,CH₂CH₂CH₂F, C₁₋₆ alkyl, C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl, with theproviso that the compound excludesN-(((2R,3R,4R,5R)-3,4-dihydroxy-5-methylpyrrolidin-2-yl)methyl)acetamide.2. The compound of claim 1 wherein the compound is selected from thefollowing group:N-ethyl-2-((2R,3R,4R,5S)-5-(fluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin-2-yl)acetamide;2-((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin-2-yl)-N-ethylacetamide;2-((2R,3R,4R,5S)-5-(fluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide;2-((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide;2-((2R,3R,4R,5R)-3,4-dihydroxy-5-methyl-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide;2-((2R,3R,4R,5S)-3-fluoro-5-(fluoromethyl)-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide;2-((2R,3S,4R,5S)-5-(difluoromethyl)-3-fluoro-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide;N-(((2R,3R,4R,5S)-5-(fluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)propionamide;N-(((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)propionamide;N-(((2R,3R,4R,5R)-3,4-dihydroxy-5-methyl-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)propionamide;N-(((2R,3R,4R,5S)-3-fluoro-5-(fluoromethyl)-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)propionamide;N-(((2R,3S,4R,5S)-5-(difluoromethyl)-3-fluoro-4-hydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)propionamide;or a pharmaceutically acceptable salt of any of the foregoing compounds.3. A pharmaceutical composition comprising the compound of claim 1 or apharmaceutically acceptable salt thereof in combination with apharmaceutically acceptable carrier.
 4. A pharmaceutical compositioncomprising a compound of claim 2 or a pharmaceutically acceptable saltthereof in combination with a pharmaceutically acceptable carrier. 5.The compound of claim 1 wherein the compound is selected from thefollowing group:N-ethyl-2-((2R,3R,4R,5S)-5-(fluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin-2-yl)acetamide;2-((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-phenethylpyrrolidin-2-yl)-N-ethylacetamide;2-((2R,3R,4R,5S)-5-(fluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide;2-((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)-N-methylacetamide;N-(((2R,3R,4R,5S)-5-(fluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)propionamide;N-(((2R,3R,4R,5S)-5-(difluoromethyl)-3,4-dihydroxy-1-(3-phenylpropyl)pyrrolidin-2-yl)methyl)propionamide;or a pharmaceutically acceptable salt of any of the foregoing compounds.6. A pharmaceutical composition comprising a compound of claim 5 or apharmaceutically acceptable salt thereof in combination with apharmaceutically acceptable carrier.